CN103946221B

CN103946221B - For treating the heterocyclic compound of cystic fibrosis

Info

Publication number: CN103946221B
Application number: CN201280056105.8A
Authority: CN
Inventors: K·J·巴拉; R·巴特勒; S·P·科林伍德; E·C·霍尔; L·爱德华兹; D·M·莱格兰德; K·施皮格尔
Original assignee: Novartis AG
Current assignee: Novartis AG
Priority date: 2011-09-16
Filing date: 2012-09-14
Publication date: 2016-08-03
Anticipated expiration: 2032-09-14
Also published as: CN103946221A; EP2755967B1; JP2014526500A; WO2013038386A1; ES2558457T3; JP5886433B2; US9034879B2; US20140228376A1; EP2755967A1

Abstract

The invention provides the pyridine diazole/thiadiazoles derivative of Formulas I, wherein A is N or CR^4a, described derivant is for recovering or strengthen the function of saltant type and/or wild type CFTR with treatment cystic fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease, asthma, respiratory tract infection, pulmonary carcinoma, xerostomia and keratoconjunctivitis sicca or constipation (induction of IBS, IBD, opioid).Also include the pharmaceutical composition comprising this analog derivative.X is

Description

For treating the heterocyclic compound of cystic fibrosis

Invention field

The present invention relates to pyridine-diazole/thiadiazoles derivative, their preparation method and the purposes as medicine.

Background technology

Cystic fibrosis (CF) is the sudden change in the gene by encoding Cystic Fibrosis transmembrane channel regulatory factor (CFTR) The fatal genetic disease caused, described cystic fibrosis transmembrane conductance regulator is to participate in multiple organs (including lung) Salt and the epithelium anion channel of protein kinase A (PKA)-activation of fluid transport.Major part CF sudden change reduces cell surface On CFTR number of channels (such as synthesize or process sudden change) infringement channel function (such as gate or conductance sudden change) or this two Person.There is no at present and approved be directly targeted the therapy in CFTR.The invention discloses compound, it recovers or strengthens sudden change And/or the function of the CFTR of wild type is with treatment cystic fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic Obstructive pulmonary disease, asthma, respiratory tract infection, pulmonary carcinoma, xerostomia and keratoconjunctivitis sicca or constipation (IBS, IBD, opium The induction of sample material).

Invention describes

On the one hand, the invention provides compound of formula I or its officinal salt:

Wherein:

A is N or CR^4a；

X is

R¹It is selected from H；The C optionally replaced by one or more halogen atoms₁-C₈Alkyl；C₂-C₈Thiazolinyl；C₂-C₈Alkynyl； C₃-C₁₀Cycloalkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；Optionally replaced by one or more halogen atoms C₁-C₈Alkoxyl；Halogen；SO₂NR⁸R⁹；SO₂R¹⁰；The S-C optionally replaced by one or more halogen atoms₁-C₈Alkyl；S-C₆- C₁₄Aryl；-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；With-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals；Wherein said heterocyclic radical comprises At least one is selected from the hetero atom of N, O and S；CN；NR¹¹R¹²；CONR¹³R¹⁴；NR¹³SO₂R¹⁵；NR¹³C(O)R¹⁵And CO₂R¹⁵, wherein Described cycloalkyl, cycloalkenyl group, aryl and heterocyclyl groups are the most optionally replaced by one or more Z substituent groups；

R²It is C₁-C₄Haloalkyl；

R³It is H or the C optionally replaced by one or more halogen atoms₁-C₈Alkyl；

R⁴It is H；The C optionally replaced by one or more halogen atoms₁-C₈Alkyl；C₂-C₈Thiazolinyl；C₂-C₈Alkynyl；C₃-C₁₀ Cycloalkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；The C optionally replaced by one or more halogen atoms₁-C₈Alkane Epoxide；C₁-C₄Alkoxy C₁-C₄Alkyl；C₁-C₈Hydroxy alkyl；OH；CN；Fluorine；-(CH₂)_m-NR¹⁷R¹⁸；-(C₀-C₄Alkyl)-C₆- C₁₄Aryl；-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals, wherein said heterocyclic radical comprises at least one miscellaneous former selected from N, O and S Son；Or-(C₀-C₄Alkyl)-CO₂R¹⁵, wherein said cycloalkyl, cycloalkenyl group ,-(C₀-C₄Alkyl)-C₆-C₁₄Aryl and-(C₀-C₄Alkane Base)-3 to 14 yuan of heterocyclyl groups the most optionally replace by one or more Z substituent groups；

R^4aIt is selected from H；The C optionally replaced by one or more halogen atoms₁-C₄Alkyl；C₂-C₈Thiazolinyl；-(C₀-C₄Alkane Base)-C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals；C₁-C₈Hydroxy alkyl；Halogen；-(CH₂)_m-NR¹⁷R¹⁸；-(C₀- C₄Alkyl)-CO₂R¹⁵With-(C₀-C₄Alkyl)-C (O) NR¹⁷R¹⁸；Wherein said-(C₀-C₄Alkyl)-C₆-C₁₄Aryl and-(C₀-C₄Alkane Base)-3 to 14 yuan of heterocyclyl groups the most optionally replace by one or more Z substituent groups；

R⁵It is the C optionally replaced by one or more halogen atoms₁-C₈Alkyl；C₂-C₈Thiazolinyl；C₂-C₈Alkynyl；C₃-C₁₀Ring Alkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；The C optionally replaced by one or more halogen atoms₁-C₈Alcoxyl Base；Fluorine；-(CH₂)_m-NR¹⁷R¹⁸；-(CH₂)_m-OR⁴；-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-3 to 14 yuan of heterocycles Base, wherein said heterocyclic radical comprises at least one hetero atom selected from N, O and S；Or-(C₀-C₄Alkyl)-CO₂R¹⁵, wherein said Cycloalkyl, cycloalkenyl group ,-(C₀-C₄Alkyl)-C₆-C₁₄Aryl and-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclyl groups are the most optionally Replaced by one or more Z substituent groups；Or

R³And R⁴Form oxo group (C=O) together；Or

R³And R⁵3-8 unit cycloalkyl is formed together with the carbon atom being connected with them；Or

R⁴And R⁵Form 5-8 unit cycloalkyl together with the carbon atom being connected with them or 5-8 unit comprises one or more being selected from The heteroatomic heterocyclic radical of N, O and S, wherein said ring system is optionally replaced by one or more Z substituent groups；

M is 0,1,2 or 3；

R⁸、R¹¹、R¹³And R¹⁷It is H, the C optionally replaced by one or more halogen atoms independently of one another₁-C₈Alkyl, C₃- C₁₀Cycloalkyl or-(C₁-C₄Alkyl)-C₃-C₈Cycloalkyl；

R⁹、R¹⁰、R¹²、R¹⁴、R¹⁵、R¹⁶And R¹⁸It is H independently of one another；The C optionally replaced by one or more halogen atoms₁- C₈Alkyl；C₂-C₈Thiazolinyl；C₂-C₈Alkynyl；C₃-C₁₀Cycloalkyl；C₅-C₁₀Cycloalkenyl group；-C₁-C₄Alkyl-C₃-C₈Cycloalkyl；-(C₀-C₄ Alkyl)-C₆-C₁₄Aryl；Or-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals, wherein said heterocyclic radical comprises at least one selected from N, O With the hetero atom of S, wherein said cycloalkyl, cycloalkenyl group, aryl and heterocyclyl groups are the most optionally replaced by one or more Z Base replaces；Or

R⁸And R⁹、R¹¹And R¹²、R¹³And R¹⁴And R¹⁷And R¹⁸4-14 unit is formed optionally together with the nitrogen-atoms being connected with them The heterocyclic radical replaced by one or more Z substituent groups；

Z be independently OH, aryl, O-aryl, benzyl, O-benzyl, optionally by one or more OH groups or NH₂Group takes The C in generation₁-C₆Alkyl, the C optionally replaced by one or more halogen atoms₁-C₆Alkyl, optionally by one or more OH groups or C₁-C₄The substituted C of alkoxyl₁-C₆Alkoxyl, NR³⁰(SO₂)R³²、(SO₂)NR³¹R³²、(SO₂)R³²、NR³⁰C(O)R³²、C(O) NR³¹R³²、NR³⁰C(O)NR³¹R³²、NR³⁰C(O)OR¹⁹、NR³¹R³²、C(O)OR³¹、C(O)R³¹、SR³¹、OR³¹, oxo, CN, NO₂, halogen Element or 3 to 14 yuan of heterocyclic radicals, wherein said heterocyclic radical comprises at least one hetero atom selected from N, O and S；

R³⁰It is H or C₁-C₆Alkyl；

R³¹And R³²It is H independently of one another；C₁-C₈Alkyl；C₃-C₈Cycloalkyl；C₁-C₄Alkoxy-C₁-C₄Alkyl；(C₀-C₄Alkane Base)-aryl, it optionally replaces selected from following group by one or more: C₁-C₆Alkyl, C₁-C₆Alkoxyl and halogen； (C₀-C₄Alkyl)-3-to 14-unit heterocyclic radical, described heterocyclic radical comprises one or more hetero atom selected from N, O and S, and it is optional Ground is replaced selected from following group by one or more: halogen, oxo, C₁-C₆Alkyl and C (O) C₁-C₆Alkyl；(C₀-C₄Alkane Base)-O-aryl, it optionally replaces selected from following group by one or more: C₁-C₆Alkyl, C₁-C₆Alkoxyl and halogen； (C₀-C₄Alkyl)-O-3-to 14-unit heterocyclic radical, described heterocyclic radical comprises one or more hetero atom selected from N, O and S, its Optionally replaced selected from following group by one or more: halogen, C₁-C₆Alkyl or C (O) C₁-C₆Alkyl；Wherein said alkane Base group is optionally replaced by one or more following groups: halogen atom, C₁-C₄Alkoxyl, C (O) NH₂、C(O)NHC₁-C₆Alkane Base or C (O) N (C₁-C₆Alkyl)₂；Or

R³¹And R³²Forming 5-to 10-unit heterocyclic radical together with the nitrogen-atoms being connected with them, described heterocyclic radical comprises one Or multiple other hetero atom selected from N, O and S, this heterocyclic radical is optionally replaced selected from following substituent group by one or more: OH；Halogen；Aryl；5-to 10-unit heterocyclic radical, it comprises one or more hetero atom selected from N, O and S；S(O)₂-aryl；S (O)₂-C₁-C₆Alkyl；The C optionally replaced by one or more halogen atoms₁-C₆Alkyl；Optionally by one or more OH groups or C₁-C₄The substituted C of alkoxyl₁-C₆Alkoxyl；With C (O) OC₁-C₆Alkyl, wherein said aryl and heterocyclyl substituent self are appointed Selection of land is by C₁-C₆Alkyl, C₁-C₆Haloalkyl or C₁-C₆Alkoxyl replaces.

This document describes the multiple embodiments of the present invention.It is believed that the spy described in detail in each embodiment Levy the feature that can describe in detail with other and be combined to provide other embodiment.

In one embodiment of the present invention as described in the most Anywhere, wherein A is CR^4aAnd R^4aBe selected from halogen, The C optionally replaced by one or more halogen atoms₁-C₄Alkyl；C₂-C₈Thiazolinyl and-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；Wherein- (C₀-C₄Alkyl)-C₆-C₁₄Aryl can optionally be replaced by one or more Z substituent groups.

In one embodiment of the present invention as described in the most Anywhere, wherein A is CR^4aAnd R^4aBe selected from halogen, The C optionally replaced by one or more halogen atoms₁-C₄Alkyl；C₂-C₈Thiazolinyl and-(C₀-C₄Alkyl)-C₆-C₁₄Aryl.

In one embodiment of the present invention as described in the most Anywhere, wherein A is CR^4aAnd R^4aIt is selected from chlorine, second Base, isopropyl, isopropenyl and phenyl；Wherein phenyl can optionally be replaced by one or more Z substituent groups.

In one embodiment of the present invention as described in the most Anywhere, wherein X is

In one embodiment of the present invention as described in the most Anywhere, wherein

R¹It is selected from H；The C optionally replaced by one or more halogen atoms₁-C₈Alkyl；C₃-C₁₀Cycloalkyl；Optionally by one Individual or multiple substituted C of halogen atom₁-C₈Alkoxyl；Halogen；C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals its Described in heterocyclic radical comprise at least one hetero atom selected from N, O and S；And NR¹¹R¹², wherein said aryl and heterocyclic radical are each Optionally replaced by one or more Z substituent groups.

In one embodiment of the present invention as described in the most Anywhere, wherein R¹Be selected from H, optionally by one or The substituted C of multiple halogen atoms₁-C₄Alkyl；C₃-C₁₀Cycloalkyl；The C optionally replaced by one or more halogen atoms₁-C₄ Alkoxyl, and halogen.

In one embodiment of the present invention as described in the most Anywhere, wherein R¹It is selected from optionally by one or many The substituted C of individual halogen atom₁-C₄Alkyl；C₃-C₁₀Cycloalkyl；The C optionally replaced by one or more halogen atoms₁-C₄Alkane Epoxide, and halogen.

R¹It is selected from H, methoxyl group, trifluoromethyl, bromine, cyclopropyl and methyl.

R¹Being aryl, wherein aryl is the phenyl optionally replaced by one or more Z substituent groups.

R²It is CF₃。

R³It is selected from H or the C that optionally replaced by one or more halogen atoms₁-C₄Alkyl.

In one embodiment of the present invention as described in the most Anywhere, wherein R⁴It is selected from H；Optionally by one or The substituted C of multiple halogen atoms₁-C₄Alkyl；The C optionally replaced by one or more halogen atoms₁-C₄Alkoxyl；- (CH₂)_m-NR¹⁷R¹⁸And OH；R¹⁷And R¹⁸It is H independently of one another；The C optionally replaced by one or more halogen atoms₁-C₈Alkyl.

In one embodiment of the present invention as described in the most Anywhere, wherein R⁵It is selected from H；Optionally by one or The substituted C of multiple halogen atoms₁-C₄Alkyl；The C optionally replaced by one or more halogen atoms₁-C₄Alkoxyl；OH；CN； Halogen；-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；With-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals；Wherein said heterocyclic radical comprise to A few hetero atom selected from N, O and S, wherein aryl and heterocyclyl groups are the most optionally taken by one or more Z substituent groups Generation.

In one embodiment of the present invention as described in the most Anywhere, wherein R³And R⁵The carbon being connected with them Atom forms 3-6 unit cycloalkyl together.

In one embodiment of the present invention as described in the most Anywhere, wherein R⁴And R⁵The carbon being connected with them Atom forms 5-6 unit cycloalkyl together or 5-6 unit comprises one or more heteroatomic heterocyclic radical selected from N, O and S, the most miscellaneous Ring group is optionally replaced by one or more Z substituent groups.

R³It is selected from H or optionally by one and the C of multiple halogen atoms replacement₁-C₄Alkyl；

R⁴It is selected from H；The C optionally replaced by one or more halogen atoms₁-C₄Alkyl；Optionally by one or more halogen The element substituted C of atom₁-C₄Alkoxyl；-(CH₂)_m-NR¹⁷R¹⁸And OH；

R⁵It is selected from H；The C optionally replaced by one or more halogen atoms₁-C₄Alkyl；Optionally by one or more halogen The element substituted C of atom₁-C₄Alkoxyl；OH；CN；Halogen；-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；With-(C₀-C₄Alkyl)-3 to 14 Unit's heterocyclic radical；Wherein said heterocyclic radical comprises at least one hetero atom selected from N, O and S, and wherein aryl and heterocyclyl groups are each From optionally being replaced by one or more Z substituent groups；Or

R³And R⁵3-6 unit cycloalkyl is formed together with the carbon atom being connected with them；Or

R⁴And R⁵Form 5-6 unit cycloalkyl together with the carbon atom being connected with them or 5-6 unit comprises one or more being selected from The heteroatomic heterocyclic radical of N, O and S, wherein heterocyclic radical is optionally replaced by one or more Z substituent groups；

R¹⁷And R¹⁸It is H independently of one another；The C optionally replaced by one or more halogen atoms₁-C₈Alkyl.

A is CR^4a；

X is

R¹It is selected from H；The C optionally replaced by one or more halogen atoms₁-C₄Alkoxyl；Optionally by one or The substituted C of multiple halogen atoms₁-C₄Alkoxyl；

R²It is CF₃；

R³It is H, CH₃Or CF₃；

R⁴It is H or Me；

R⁵It is phenyl ,-NR¹⁷R¹⁸Or OH；And

A is CR^4a；

X is

R¹It is selected from the C optionally replaced by one or more halogen atoms₁-C₄Alkoxyl；Optionally by one or many The substituted C of individual halogen atom₁-C₄Alkoxyl；

R²It is CF₃；

R³It is H, CH₃Or CF₃；

R⁴It is H or Me；

R⁵It is-NR¹⁷R¹⁸Or OH；And

In one embodiment of the present invention as described in the most Anywhere, compound of formula I comprise Formula II compound or Its officinal salt:

Wherein A, R¹、R²、R³And R^4aAs defined in embodiment 1-16；And

R¹⁰¹It is selected from following group:

In one embodiment of the present invention as described in the most Anywhere, wherein A is CR^4a, wherein R^4aIt is H.

In one embodiment of the present invention as described in the most Anywhere, wherein A is CR^4a；

R¹It is the C optionally replaced by one or more halogen atoms₁-C₄Alkyl；

R¹⁰¹It is

In one embodiment of the present invention as described in the most Anywhere, wherein R¹⁰¹It is

In one embodiment of the present invention as described in the most Anywhere, A is N.

In one embodiment of the present invention as described in the most Anywhere, A is CR^4a, wherein R^4aIt is H or optional quilt The substituted C of one or more halogen atoms₁-C₄Alkyl；

In one embodiment of the present invention as described in the most Anywhere, A is CR^4a, wherein R^4aIt is H, methyl or second Base.

In one embodiment of the present invention as described in the most Anywhere, A is CR^4a, wherein R^4aIt is H.

In one embodiment of the present invention as described in the most Anywhere, X is

Embodiment of the present invention as defined above provide compound of formula I, wherein

A is CR^4a；

X is

R¹It is halogen, the C optionally replaced by one or more halogen atoms₁-C₄Alkyl or optionally by one or more halogen The element substituted C of atom₁-C₄Alkoxyl；

R²It is C₁-C₄Haloalkyl；

R³It is H or Me；

R^4aIt is H；

R⁴It is-(CH₂)_m-NR¹⁷R¹⁸；-(CH₂)_m-OR³；Or OH；

M is 0,1 or 2；

R⁵It is the C optionally replaced by one or more halogen atoms₁-C₄Alkyl；Optionally by one or more halogen atoms Substituted C₁-C₄Alkoxyl；-(CH₂)_m-NR¹⁷R¹⁸；-(CH₂)_m-OR⁴；Or-(C₀-C₄Alkyl)-C₆-C₁₀Aryl, wherein-(C₀-C₄ Alkyl)-C₆-C₁₀Aryl is optionally replaced by one or more Z substituent groups；Or

R⁴And R⁵Form 5-6 unit together with the carbon atom being connected with them and comprise one or more miscellaneous former selected from N, O and S The heterocycle ring system of son, wherein said ring system is optionally replaced by one or more Z substituent groups；And

R¹⁷And R¹⁸It is H independently of one another；The C optionally replaced by one or more halogen atoms₁-C₄Alkyl.

A is CR^4a；

X is

R¹It is the C optionally replaced by one or more halogen atoms₁-C₄Alkyl；

R²It is C₁-C₄Haloalkyl；

R³It is H or the C optionally replaced by one or more halogen atoms₁-C₄Alkyl；

R^4aIt is H；

R⁴It is-(CH₂)_m-NR¹⁷R¹⁸；-(CH₂)_m-OR³；Or OH；

M is 0,1 or 2；

R⁵It is the C optionally replaced by one or more halogen atoms₁-C₄Alkyl；Optionally by one or more halogen atoms Substituted C₁-C₄Alkoxyl；-(CH₂)_m-NR¹⁷R¹⁸；-(CH₂)_m-OR⁴；Or C₆-C₁₀Aryl, wherein C₆-C₁₀Aryl is optionally by one Individual or multiple Z substituent groups replace；Or

A is CR^4a；

X is

R¹It is the C optionally replaced by one or more halogen atoms₁-C₄Alkoxyl；

R²It is C₁-C₄Haloalkyl；

R^4aIt is H；

R⁴It is-(CH₂)_m-NR¹⁷R¹⁸；-(CH₂)_m-OR³；Or OH；

It is 0,1 or 2；

R⁵It is the C optionally replaced by one or more halogen atoms₁-C₄Alkyl；Optionally by one or more halogen atoms Substituted C₁-C₄Alkoxyl；Or-C₆-C₁₀Aryl, wherein C₆-C₁₀Aryl is optionally replaced by one or more Z substituent groups；Or

A is CR^4a；

X is

R²It is C₁-C₄Haloalkyl；

R³It is H, methyl or trifluoromethyl；

R^4aIt is H；

R⁴It is-NR¹⁷R¹⁸；Or OH；

R⁵It is the C optionally replaced by one or more halogen atoms₁-C₄Alkyl；Phenyl, wherein said phenyl optionally by One or more Z substituent groups replace；And

A is CR^4a；

X is

R¹It is the C optionally replaced by one or more halogen atoms₁-C₄Alkyl；

R²It is C₁-C₄Haloalkyl；

R³It is H, methyl or trifluoromethyl；

R^4aIt is H；

R⁴It is-NR¹⁷R¹⁸；Or OH；

A is CR^4a；

X is

R²It is C₁-C₄Haloalkyl；

R³It is H, methyl or trifluoromethyl；

R^4aIt is H；

R⁴It is-NR¹⁷R¹⁸；Or OH；

R⁵It is the C optionally replaced by one or more halogen atoms₁-C₄Alkyl；And

The another embodiment of the present invention as defined above provides that to have the purest mapping of R configuration different The compound of structure body.

The another embodiment of the present invention as defined above provides that to have the purest mapping of S configuration different The compound of structure body.

Some compound of formula I comprises Formula II compound or pharmaceutically acceptable salt thereof,

Wherein A, R¹、R²、R³And R^4aThere is the definition of Formulas I；And

R¹⁰¹It is selected from following group:

In the another embodiment of the Formula II of the present invention, A is CR^4a, wherein R^4aIt is H.

In the another embodiment of the Formula II of the present invention, R¹It is selected from H；Optionally taken by one or more halogen atoms The C in generation₁-C₄Alkyl；The C optionally replaced by one or more halogen atoms₁-C₄Alkoxyl；Halogen；C₆-C₁₄Aryl；-(C₀- C₄Alkyl)-3 to 14 yuan of heterocyclic radicals, wherein said heterocyclic radical comprises at least one hetero atom selected from N, O and S；And NR¹¹R¹², its Described in aryl and heterocyclic radical the most optionally replaced by one or more Z substituent groups.

In the another embodiment of the Formula II of the present invention, R¹It is the C optionally replaced by one or more halogen atoms₁- C₄Alkyl, the C optionally replaced by one or more halogen atoms₁-C₄Alkoxyl；Halogen；C₆Aryl；Or hexa-member heterocycle base, its Described in heterocyclic radical comprise at least one hetero atom selected from N, O and S, wherein said aryl and heterocyclic radical are the most optionally by one Individual or multiple Z substituent groups replace.

In the another embodiment of the Formula II of the present invention, R¹It is the C optionally replaced by one or more halogen atoms₁- C₄Alkyl, the C optionally replaced by one or more halogen atoms₁-C₄Alkoxyl；Or halogen.

In the another embodiment of the Formula II of the present invention, R³It is H or methyl.

In the another embodiment of the Formula II of the present invention, R^4aIt is H

In one embodiment of the Formula II of the present invention as described in the most Anywhere, X is

Embodiment of the present invention as defined above provide Formula II compound, wherein

A is CR^4a；

R¹It it is halogen；

R^4aIt is H；

R¹⁰¹It is

A is CR^4a；

R¹It is the C optionally replaced by one or more halogen atoms₁-C₄Alkyl；

R^4aIt is H；

R¹⁰¹It is

CR^4a；

R^4aIt is H；

R¹⁰¹It is

A is CR^4a；

R^4aIt is H；

R¹⁰¹It is

A is CR^4a；

R^4aIt is H；

R¹⁰¹It is

A is CR^4a；

R^4aIt is H；

R¹⁰¹It is

The another embodiment of the present invention as defined above provides Formulas I and Formula II compound or it is pharmaceutically acceptable Salt, is represented by following compound:

2-(5-(3-amino-6-methoxyl group-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1, 1-trifluoro propan-2-ol (racemic)；

(R)-2-(5-(3-amino-6-methoxyl group-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)- 1,1,1-trifluoro propan-2-ol；

(S)-2-(5-(3-amino-6-methoxyl group-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)- 1,1,1-trifluoro propan-2-ol；

3-(5-benzyl-1,3,4-diazole-2-base)-5-bromo-6-(trifluoromethyl) pyrazine-2-amine；

(5-(3-amino-6-bromo-5-(trifluoromethyl) pyrazine-2-base)-1,3,4-diazole-2-base) (phenyl) ketone；

2-(5-(3-amino-5,6-two (trifluoromethyl) pyrazine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoro Propan-2-ol；

2-(5-(3-amino-5,6-two (trifluoromethyl) pyrazine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoro Propan-2-ol (racemic)；

(R)-2-(5-(3-amino-5,6-two (trifluoromethyl) pyrazine-2-base)-1,3,4-diazole-2-base)-1,1,1- Trifluoro propan-2-ol；

(S)-2-(5-(3-amino-5,6-two (trifluoromethyl) pyrazine-2-base)-1,3,4-diazole-2-base)-1,1,1- Trifluoro propan-2-ol；

2-(5-benzyl-1,3,4-diazole-2-base)-6-bromo-5-(trifluoromethyl) pyridine-3-amine；

2-(5-benzyl-[1,3,4] diazole-2-base)-5-trifluoromethylpyridin-3-base-amine；

2-[5-(the fluoro-benzyl of 4-)-[1,3,4] diazole-2-base]-5-trifluoromethylpyridin-3-base-amine；

The bromo-2-of 6-[5-(the fluoro-benzyl of 4-)-[1,3,4] diazole-2-base]-5-trifluoromethylpyridin-3-base amine；

The bromo-2-of 6-[5-(2,2,2-trifluoro ethyl)-[1,3,4] diazole-2-base]-5-trifluoromethylpyridin-3-base Amine；

2-(5-(3-amino-6-cyclopropyl-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1, 1-trifluoro propan-2-ol；

2-(5-(3-amino-6-methyl-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1- Trifluoro propan-2-ol；

2-(5-(3-amino-6-bromo-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-three Fluorine propan-2-ol；

2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoropropyl- 2-alcohol；

(2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoropropyl- 2-alcohol (racemic).

(R)-2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoro Propan-2-ol；

(S)-2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoro Propan-2-ol；

2-(5-(3-amino-6-bromo-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base) propan-2-ol；

(R)-2-(5-(amino (phenyl) methyl)-1,3,4-diazole-2-base)-6-bromo-5-(trifluoromethyl) pyridine-3- Amine；With

2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4-thiadiazoles-2-base)-1,1,1-trifluoropropyl- 2-alcohol.

(R)-2-[5-(3-amino-4-chloro-6-methoxyl group-5-trifluoromethylpyridin-2-base)-[1,3,4] diazole-2- Base]-1,1,1-trifluoro-propyl-2-alcohol；

(S)-2-(5-(3-amino-4-chloro-6-methoxyl group-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2- Base)-1,1,1-trifluoro propan-2-ol；

(S)-2-[5-(3-amino-4-ethyl-6-methoxyl group-5-trifluoromethylpyridin-2-base)-[1,3,4] diazole- 2-yl]-1,1,1-trifluoro-propyl-2-alcohol；

(S)-2-[5-(3-amino-4-isopropenyl-6-methoxyl group-5-trifluoromethylpyridin-2-base)-[1,3,4] Diazole-2-base]-1,1,1-trifluoro-propyl-2-alcohol；

(S)-2-[5-(3-amino-4-isopropyl-6-methoxyl group-5-trifluoromethylpyridin-2-base)-[1,3,4] two Azoles-2-base]-1,1,1-trifluoro-propyl-2-alcohol；With

(S)-2-[5-(3-amino-6-methoxyl group-4-phenyl-5-trifluoromethylpyridin-2-base)-[1,3,4] diazole- 2-yl]-1,1,1-trifluoro-propyl-2-alcohol.

It should be appreciated that any and all embodiment of the present invention can combine with arbitrarily other embodiment to retouch State the other embodiments of the present invention.Additionally, any key element of an embodiment can with from any embodiment Arbitrarily with other factor combinations all to describe other embodiments.It will be apparent to a skilled person that can not Substituent group combination in the case of energy is not the aspect of the present invention.

The particularly preferred particular compound of formula (I) or Formula II is those described in embodiments below compounds.

Definition

The term used in this specification has a following implication:

" optionally it is replaced " and means in the group that described group can be listed on one or more positions subsequently Any one or its any combination are replaced.

" optionally replaced " the relevant group of expression by one or more Z group and can include one or more substituent group, described Substituent group is each independently selected from the definition of Z included group.Therefore, when there being two or more Z group substituent groups, They can be identical or different.

" halo " used herein or " halogen " can be fluorine, chlorine, bromine or iodine.

" C used herein₁-C₈-alkyl " represent the straight or branched alkyl with 1-8 carbon atom.If providing difference The carbon atom of quantity, such as C₆Or C₃, then this definition should correspondingly be modified, such as " C₁-C₄-alkyl " represent methyl, second Base, propyl group, isopropyl, butyl, isobutyl group, sec-butyl and the tert-butyl group.

" C used herein₁-C₈-alkoxyl " represent the alkoxyl of straight or branched with 1-8 carbon atom.If given Go out the carbon atom of varying number, such as C₆Or C₃, then this definition should correspondingly be modified, such as " C₁-C₄-alkoxyl " represent Methoxyl group, ethyoxyl, propoxyl group, isopropoxy, butoxy, isobutoxy, sec-butoxy and tert-butoxy.

" C used herein₁-C₄-haloalkyl " represent at least one of which hydrogen by halogen replace to have 1-4 carbon former The alkyl of the straight or branched of son.If providing the carbon atom of varying number, such as C₆Or C₃, then this definition should correspondingly be carried out Amendment, such as " C₁-C₄-haloalkyl " represent methyl that at least one hydrogen replaced by halogen, ethyl, propyl group, isopropyl, butyl, Isobutyl group, sec-butyl and the tert-butyl group, when wherein halogen is fluorine: (CF₃)₂CH-、CH₃-CF₂-、CF₃CF₂-、CF₃、CF₂H-、 CF₃CF₂CHCF₃Or CF₃CF₂CF₂CF₂-。

" C used herein₁-C₈-hydroxy alkyl " represent the straight or branched alkyl with 1-8 carbon atom, at least a part of which One hydrogen is replaced by oh group.If different carbon numbers is designated (such as C₆Or C₃), then this definition is repaiied accordingly Just, such as " C₁-C₄-hydroxy alkyl " represent at least one of which hydrogen replaced by oh group methyl, ethyl, propyl group, isopropyl Base, butyl, isobutyl group, sec-butyl and the tert-butyl group.

Term " C used herein_2-8Thiazolinyl " refer to comprise straight containing at least one carbon-carbon double bond of 2-8 carbon atom Chain or the unsaturated alkyl of side chain.The example of this type of group includes vinyl, acrylic, cyclobutenyl and pentenyl.Unless specified Specific structure, term cyclobutenyl and pentenyl etc. comprise the most possible E and Z isomer.

Term " C used herein_3-8Cycloalkyl " refer to the saturated monocycle hydrocarbon ring of 3-6 carbon atom.The example of this type of group Including cyclopropyl, cyclobutyl, cyclopenta and cyclohexyl.

Term " alkylidene " represents the straight or branched saturated hydrocarbon chain comprising 1-8 carbon atom.If different carbon atoms Number is designated (such as C₆Or C₃), then this definition is modified accordingly.

" amino-C₁-C₈-alkyl " and " amino-C₁-C₈-alkoxyl " represent that amino is by nitrogen-atoms and C₁-C₈-alkyl is even Connect, such as NH₂-(C₁-C₈)-, or and C₁-C₈-alkoxyl connects, such as NH₂-(C₁-C₈)-O-.If different carbon numbers It is designated (such as C₆Or C₃), then this definition is modified accordingly.

“C₁-C₈-alkyl amino " and " two (C₁-C₈-alkyl) amino " represent C as defined hereinabove₁-C₈-alkyl, it leads to Cross carbon atom to be connected with amino group.At two (C₁-C₈-alkyl) described C in amino₁-C₈-alkyl group can be identical or not With.If different carbon numbers is designated (such as C₆Or C₃), then this definition is modified accordingly.

" amino-(hydroxyl)-C₁-C₈-alkyl " represent that amino is by nitrogen-atoms and C₁-C₈-alkyl connects, and hydroxyl passes through Oxygen atom and identical C₁-C₈-alkyl connects.If different carbon numbers is designated (such as C₆Or C₃), then this definition phase Should be modified.

" C used herein₁-C₈-alkyl-carbonyl " and " C₁-C₈-alkoxy carbonyl " represent C as defined above₁-C₈- Alkyl or C₁-C₈-alkoxyl is connected with carbonyl group by carbon atom respectively.If different carbon numbers is designated (such as C₆ Or C₃), then this definition is modified accordingly.

" C used herein₃-C₈-naphthene base carbonyl " represent C as defined above₃-C₈-cycloalkyl by carbon atom with Carbonyl group connects.If different carbon numbers is designated (such as C₆Or C₃), then this definition is modified accordingly.

" C used herein₇-C₁₄-aralkyl " represent alkyl as defined above (such as C₁-C₄-alkyl) by as herein Defined C₆-C₁₀-aromatic carbocyclic radical replaces.If different carbon numbers is designated (such as C₆Or C₃), then this definition It is modified accordingly.

" C used herein₃-C₁₅-cycloalkyl " represent the ring with the saturated of 3-15 ring carbon atom or fractional saturation Alkyl, such as C₃-C₈-cycloalkyl.C₃-C₁₅The example of-cycloalkyl includes but not limited to cyclopropyl, cyclobutyl, cyclopenta, hexamethylene Base, suberyl or ring octyl group or bicyclic groups, such as bicyclooctyl, diazabicyclononyl, including indanyl and indenyl and bicyclo-decyl. If providing the carbon atom of varying number, such as C₆, then this definition should correspondingly be modified.

" aryl " used herein or " C₆-C₁₅-aromatic carbocyclyl groups " represent the aromatic group with 6-15 ring carbon atom Group.C₆-C₁₅The example of-aromatic carbocyclic radical includes but not limited to phenyl, phenylene, benzene three base, naphthyl, naphthylene, naphthalene three base Or anthrylene.If providing the carbon atom of varying number, such as C₁₀, then this definition should correspondingly be modified.

" 4-8 unit heterocyclic radical ", " 5-6 unit heterocyclic radical ", " 3-10 unit heterocyclic radical ", " 3-14 unit heterocyclic radical ", " 4-14 Unit's heterocyclic radical " and " 5-14 unit heterocyclic radical " refer to respectively first selected from the 4-8 of nitrogen, the ring hetero atom of oxygen and sulfur containing at least one, 5-6 unit, 3-10 unit, 3-14 unit, 4-14 unit and the heterocycle of 5-14 unit, it can be saturated, fractional saturation or insatiable hunger (aromatics) of sum.Heterocyclic group includes monocyclic groups, condensed ring group and bridged group.The example of heterocyclic group includes but does not limits In furan, pyrroles, pyrrolidine, pyrazoles, imidazoles, triazole, different triazole, tetrazolium, thiadiazoles, isothiazole,Diazole, pyridine, piperazine Pyridine, pyrazine,Azoles, differentAzoles, pyrazine, pyridazine, pyrimidine, piperazine, pyrrolidine, ketopyrrolidine, morpholine, triazine,Piperazine, four Hydrogen furan, Tetramethylene sulfide, tetrahydric thiapyran, Pentamethylene oxide., 1,4-bis-Alkane, 1,4-oxa-thia hexamethylene, indazole, quinoline, Yin Azoles, indole, 8-aza-bicyclo [3.2.1] octane or thiazole.

A second aspect of the present invention provides as Formulas I or the compound of II defined in this paper arbitrary portion of medicine.

Another aspect of the present invention provides for treating inflammatory or allergic conditions, particularly inflammatory or obstructive gas Tract disease or the Formulas I of mucosal hydration (mucosal hydration) or the compound of II.Described disease includes such as capsule fiber Change, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease, asthma, respiratory tract infection, pulmonary carcinoma, dry mouth Disease and keratoconjunctivitis sicca or constipation (induction of IBS, IBD, opioid).

Another aspect of the present invention provides the free form defined in any one of the embodiment above or can The formula (I) of acceptable salt or the compound of (II) purposes in preparing medicine, described medicine is used for treating inflammatory or allergic effect Sexually transmitted disease (STD) disease, particularly inflammatory or obstructive airway diseases or mucosal hydration.

One embodiment of the invention provide the free form defined in any one of the embodiment above or The formula (I) of pharmaceutical acceptable salt or the compound of (II) purposes in preparing medicine, described medicine is selected from capsule for treatment Fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease, asthma, respiratory tract infection, pulmonary carcinoma, mouth Xerosis and keratoconjunctivitis sicca or the inflammatory of constipation (induction of IBS, IBD, opioid) or allergic conditions.

One embodiment of the invention provides as any one of embodiments hereinbefore defined free or pharmaceutically acceptable The formula (I) of salt form or (II) compound are used for the purposes treating in the medicine of inflammatory or allergic conditions, Qi Zhongsuo in preparation Stating disease is cystic fibrosis.

One embodiment of the invention provides disease or the method for disease for preventing or treat CFTR mediation, its Including at least one compound as herein described using effective dose to the individuality needing this kind for the treatment of.Described CFTR mediation Disease or disease selected from cystic fibrosis, primary ciliary dyskinesia, chronic bronchitis, chronic obstructive pulmonary disease, asthma, Respiratory tract infection, pulmonary carcinoma, xerostomia and keratoconjunctivitis sicca or constipation (induction of IBS, IBD, opioid).

Unless the context otherwise requires, otherwise in this specification and subsequent claim, term " comprises " or it Version such as " includes " being understood to refer to include described entirety or step or the overall or group of step, but does not arranges Except any other entirety or step or the overall or group of step.

Term used herein " officinal salt " refers to remain the biological effectiveness of the compound of the present invention and characteristic Salt, it is generally in terms of biology or other side does not have undesirable character.In some cases, the compound of the present invention Can be by there is amino and/or carboxyl or group similar therewith formation hydrochlorate and/or alkali salt.

Pharmaceutically useful acid-addition salts can be formed with mineral acid and organic acid, such as acetate, aspartate, benzoic acid Salt, benzene sulfonate, bromide/hydrobromate, bicarbonate/carbonate, disulfate/sulfate, camsilate, chlorination Thing/hydrochlorate, chloro theophylline salt (chlortheophyllonate), citrate, ethanedisulphonate, fumarate, glucoheptonic acid Salt, gluconate, glucuronate, hippurate, hydriodate/iodide, isethionate, lactate, Lactobionate, Lauryl sulfate, malate, maleate, malonate, mandelate, mesylate, Methylsulfate, naphthoic acid Salt, naphthalene sulfonate, nicotinate, nitrate, octadecanoate, oleate, oxalates, palmitate, embonate, phosphate/ Hydrophosphate/dihydric phosphate, Polygalacturonate, propionate, stearate, succinate, sulfosalicylate, winestone Hydrochlorate, toluene fulfonate and trifluoroacetate.

Such as hydrochloric acid, hydrobromic acid, sulphuric acid, nitric acid, phosphoric acid etc. can be included from its derivative mineral acid obtaining salt.

Can from its derivative organic acid obtaining salt include such as acetic acid, propanoic acid, glycolic, oxalic acid, maleic acid, the third two Acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid, methanesulfonic acid, ethyl sulfonic acid, toluenesulfonic acid and sulfosalisylic Acid etc..

Pharmaceutically useful base addition salts can be formed with inorganic base and organic base.

Such as ammonium salt and the metal from periodic table of elements I-XII race can be included from its derivative inorganic base obtaining salt. In certain embodiments, salt is derived from sodium, potassium, ammonium, calcium, magnesium, ferrum, silver, zinc and copper；Particularly suitable salt include ammonium, potassium, Sodium, calcium and magnesium salt.

Can derive from it and obtain the amine that the organic base of salt includes such as primary, secondary and tertiary amine, is replaced, including naturally occurring The amine being replaced, cyclic amine, deacidite etc..Some organic amine includes isopropylamine, benzyl star salt (benzathine), choline hydrochlorate (cholinate), diethanolamine, diethylamine, lysine, meglumine, piperazine and amino fourth three Alcohol.

The officinal salt of the present invention can be closed by alkalescence or the acidic moiety of parent compound by conventional chemical method Become.It is said that in general, can be by making the free acid form of these compounds and stoichiometric applicable alkali (such as Na, Ca, Mg Or the hydroxide of K, carbonate, bicarbonate etc.) react or by making free alkali form and the stoichiometry of these compounds Be suitable for acid reaction prepare this kind of salt.This kind of reaction is typically in water or in organic solvent or at both mixture In carry out.If it is said that in general, practical, then non-aqueous media such as ether, ethyl acetate, ethanol, isopropanol or acetonitrile Use be desirable.The list of the salt being suitable for additionally can be at such as " Remington's Pharmaceutical Sciences ", the 20th edition, Mack Publishing Company, Easton, Pa., (1985)；With Stahl's and Wermuth " Handbook of Pharmaceutical Salts:Properties, Selection and Use " (Wiley-VCH, Weinheim, Germany, 2002) in find.

Additionally, the compound of the present invention (including their salt) can also obtain with the form of its hydrate or include for Other solvent of its crystallization.

The compound of the present invention, i.e. contain the chemical combination of the formula (I) of the group that can work as hydrogen bond donor and/or receptor Thing can form eutectic together with applicable eutectic (co-crystal) forming agent.Operation can be formed by formula by known eutectic (I) compound prepares these eutectics.This generic operation includes grinding, heats, distillation, congruent melting or make formula under crystallization condition altogether (I) eutectic that compound contacts eutectic forming agent in the solution and separation is consequently formed.The eutectic forming agent being suitable for includes Those described in WO2004/078163.Therefore, present invention also offers the eutectic of the compound comprising formula (I).

Term used herein " isomer " refers to have same molecular formula but atomic arrangement different chemical combination different with configuration Thing.Additionally, the given compound that term used herein " optical isomer " or " stereoisomer " refer to the present invention can exist Various stereoisomeric configurations in any one and include geometric isomer.It should be appreciated that substituent group can be connected to On the chiral centre of carbon atom.Therefore, the present invention includes the enantiomer of described compound, diastereomer or raceme Thing." enantiomer " is a pair stereoisomer of the most stackable mirror image.The 1:1 mixture of a pair enantiomer is " raceme " mixture.In a suitable manner, this term is used for referring to racemic mixture." diastereomer " be have to But the stereoisomer of few two asymmetric atom mirror images the most each other.Absolute stereochemical is according to Cahn-lngold- Prelog R-S system specifies.When a kind of compound is pure enantiomer, the spatial chemistry in each chiral carbon can To illustrate with R or S.The compound that its absolute configuration split is not clear can be according to they Plane of rotations under sodium D-line wavelength The direction of polarized light (dextrorotation-or left-handed-) and be designated as (+) or (-).Some compound as herein described contains one or many Individual asymmetric center or axle, therefore can produce enantiomer, diastereomer and can be defined as on absolute stereochemical (R)-or (S)-other stereoisomeric forms in any ratio.The present invention includes the isomer that all such is possible, including racemic mixture, Optically pure form and intermediate blend.(R) of optically active-and (S)-isomer can be with chiral synthon or chiral reagents Prepare or split with routine techniques.If compound contains double bond, then substituent group can be E or Z configuration.If chemical combination Thing contains dibasic cycloalkyl, then naphthenic substituent can have cis-or trans-configuration.Also include all tautomerisms Form.

Any asymmetric carbon atom (such as carbon etc.) on the compound of the present invention can be enriched with raceme or enantiomer Form, such as (R)-, (S)-or (R, S)-configuration exist.In certain embodiments, each asymmetric atom at (R)-or (S)-configuration has at least 50% enantiomeric excess, at least 60% enantiomeric excess, at least 70% enantiomeric excess, at least 80% enantiomeric excess, at least 90% enantiomeric excess, at least 95% enantiomeric excess or at least 99% enantiomeric excess.If May, the substituent group on the atom with unsaturated bond can exist with cis-(Z)-or trans-(E)-form.

Therefore, the compound of the present invention used herein can be that possible isomer, rotamer, resistance turn isomery The form of one of body, tautomer or its mixture, such as, for the purest geometry (cis or trans) isomer, non- Enantiomer, optical isomer (enantiomer), racemate or its mixture.

The isomer mixture of any gained can be separated into pure or the purest according to the physics and chemistry difference of component Geometric isomer or optical isomer, diastereomer, racemate, such as, divided by chromatography and/or fractional crystallization From.

The end-product of any gained or the racemate of intermediate can be split into optical antimer, example with known method Such as its diastereoisomeric salt obtained by the acid of separation optically active or alkali the acidity or the alkali compounds that discharge optically active Split.Especially, therefore basic moiety can be used for the compound of the present invention is split into its optical antimer, such as, pass through The acid of fractional crystallization optically active such as tartaric acid, dibenzoyl tartaric acid, acetyl tartaric acid, two-O, O'-is p- The salt that toluoyl tartaric acid, mandelic acid, malic acid or Camphora-10-sulfonic acid are formed splits.Racemic product can also pass through hands Property chromatography, the high pressure lipuid chromatography (HPLC) (HPLC) of chiral sorbent is such as used to split.

Owing to the compound of the present invention is intended to be used in pharmaceutical composition, so it can be readily appreciated that each of which is preferred There is provided in a substantially pure form, for example, at least 60% pure, more suitably at least 75% pure and mild preferably at least 85%, the most extremely Few 98% pure (% is w/w).The impure prepared product of compound may be used for preparing be used in pharmaceutical composition purer Form；The prepared product of the compound that these purity are relatively low should containing at least 1%, more suitably at least 5%, preferred 10-59% The compound of the present invention.

The compound of the present invention is in a free form or its salt obtains.

When not only having there is basic group in same a part, but also when there is acidic-group, the compound of the present invention can also shape Become inner salt, such as amphion molecule.

Arbitrary structures formula given herein also aims to represent the unlabelled form of described compound and isotope labelling Form.Isotope-labeled compound has the structure that structural formula given herein is described, the most one or more Atom is replaced by the atom with selected atomic mass or mass number.The isotopic reality of the compound of the present invention can be mixed Example includes: the isotope of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine and chlorine, is the most respectively²H、³H、¹¹C、¹³C、¹⁴C、¹⁵N、¹⁸F、³¹P、³²P、³⁵S、³⁶Cl、¹²⁵I.The present invention includes various isotope-labeled compound defined herein, such as, there is radiosiotope As³H、¹³C and¹⁴Those of C.This kind of isotope-labeled compound can be used for metabolism research and (uses¹⁴C), kinetics is ground Study carefully and (such as use²H or³H), detection or imaging technique, such as positron emission tomography art (PET) or single photon emission meter Calculation machine layer image-forming art (SPECT), including medicine or substrate tissue measure of spread, or the radiation treatment for patient.Especially Ground, studies for PET or SPECT,¹⁸The compound of F or labelling is probably particularly desirable.It is said that in general, can pass through Carry out on stream or the operation disclosed in below described embodiment and preparation example, with readily available isotope labelling Reagent replace nonisotopically labelled reagent and prepare the compound of the isotope-labeled present invention.

Additionally, by heavier isotope, particularly deuterium (i.e.²H or D) replace and can provide certain because of bigger metabolic stability Treat advantage, the Half-life in vivo such as increased or the volume requirements of reduction or the improvement of therapeutic index.It should be appreciated that Deuterium is considered the substituent group of compound of formula (I) or (II) in the context of this article.This kind of heavier isotope, particularly deuterium Concentration can be defined as the isotope enrichment factor.Term used herein " the isotope enrichment factor " means isotope abundance Ratio with concrete given natural abundance of isotopes.If the substituent group on the compound of the present invention is shown deuterium, then this Kind of compound have for each D-atom specified at least 3500 (52.5% deuterium mixes on each D-atom specified), At least 4000 (60% deuterium mix), at least 4500 (67.5% deuterium mixes), at least 5000 (75% deuterium mixes), at least 5500 (82.5% deuterium mixes), at least 6000 (90% deuterium mixes), at least 6333.3 (95% deuterium mixes), at least 6466.7 (97% deuteriums Mix), at least 6600 (99% deuterium mix) or the isotope enrichment factors of at least 6633.3 (99.5% deuterium mixes).

It is said that in general, can by routine techniques well known by persons skilled in the art or by with subsidiary embodiment and preparation Method that method described in example is similar, use the isotope-labeled reagent being suitable for instead before the unlabelled reagent that uses Prepare isotope-labeled formula (I) or the compound of (II).

The pharmaceutically useful solvate of the present invention includes that wherein recrystallisation solvent can be that isotope is substituted, such as D₂O、d₆- Acetone, d₆Those solvates of-DMSO.

The compounds of this invention, i.e. comprising can be as the formula (I) of the donor of hydrogen bond and/or acceptor groups or formula (II) chemical combination Thing, can form cocrystallization with applicable eutectic forming agent.These cocrystallization can pass through from formula (I) or formula (II) compound Prepared by the eutectic forming method known.These methods include grinding, heat, distillation, congruent melting altogether, or in the solution by formula (I) Or formula (II) compound brilliant forming agent together contacts under crystallization condition, and it is subsequently isolated the cocrystallization of formation.The eutectic being suitable for Forming agent is included in those described in WO2004/078163.Therefore invention further provides and comprise formula (I) or formula (II) The cocrystallization of compound.

Synthesis

It is said that in general, can be by the approach synthesis type I described in flow process 1 to 18 and embodiment or the chemical combination of (II) Thing.

When A is CR^4aAnd R^4aWhen being alkyl, aryl or heteroaryl, compound can be synthesized according to generalized flowsheet 1.

Flow process 1

When R1 is alkyl or aryl and A is N or CH, can be according to flow process 2-in-1 one-tenth compound.B(OR^x)₂Refer to boric acid or Borate coupling reagent.

Flow process 2

When R1 is alkyl amino, and A is N or CH, compound can be synthesized according to flow process 3.

Flow process 3

Buchwald condition is applicable to metal catalytic amination, and is well known by persons skilled in the art.

R1 is Methanamide and A is CH, can synthesize compound according to flow process 4.

Flow process 4

The replacement reagent such as Mo (CO) of CO (carbon monoxide converter) gas can also be used₉。

In flow process 1-4, it is possible to use palladium catalyst such as [1,1 '-two (di-t-butyl phosphino-) ferrocene] two being suitable for Palladous chloride. (II).Skilled artisan would appreciate that and can also use other palladium catalyst.

When X is diazole, compound can be synthesized according to hereafter generalized flowsheet 5.

Flow process 5

The dehydrant being suitable for is burgess reagent or triphenylphosphine/hexachlorethane.Skilled artisan would appreciate that also Other dehydrant can be used.

When X is thiadiazole compound, compound can be synthesized according to hereafter generalized flowsheet 6.

Flow process 6

Intermediate compound I can be synthesized by the method as shown in flow process 7 or flow process 8.

Flow process 7

Or

Flow process 8

This carboxylic acid (intermediate V) is that market is obtainable, or has disclosed synthetic method.

Intermediate III can synthesize according to generalized flowsheet 9.

Flow process 9

By using method known to those skilled in the art that intermediate V esterification is obtained carboxylate.

When R1 is halogen and A is CH, intermediate II can synthesize according to generalized flowsheet 10.

Flow process 10

When R1 is halogen, and A is CR^4aAnd R^4aWhen being halogen, intermediate II can synthesize according to flow process 11.

Flow process 11

When R1 is methoxyl group and A is CH, intermediate II can synthesize according to generalized flowsheet 12.

Flow process 12

When R1 is alkoxyl and A is CH, intermediate II can synthesize according to generalized flowsheet 13.

Flow process 13

When R1 is alkoxyl, and A is CR^4aAnd R^4aWhen being halogen, intermediate II can synthesize according to generalized flowsheet 14.

Flow process 14

When R1 is alkyl, aryl or heteroaryl and A is CH, intermediate II can synthesize according to generalized flowsheet 15.

Flow process 15

The palladium catalyst being suitable for using is [1,1 '-two (di-t-butyl phosphino-) ferrocene] palladium chloride (II).This area Skilled artisan understands that and can also use other palladium catalyst.

When R1 is halogen and A is nitrogen, intermediate II can synthesize according to generalized flowsheet 16.

Flow process 16

Intermediate compound IV can synthesize according to generalized flowsheet 17 or flow process 18.

Flow process 17

Or

Flow process 18

It will be apparent to a skilled person that the general route of synthesis of above-detailed shows required is risen Beginning material carries out the popular response converted.Concrete reaction condition is not provided, but they be those skilled in the art it is known that , the condition being suitable for is considered in the range of the conventional general knowledge of those skilled in the art.

Parent material is commercially available compound or known compound and can be by organic chemistry filed Prepared by described operation.

With usual manner understood by one of ordinary skill in the art, the formula (I) of free form or the compound of (II) can be turned Chemical conversion salt form, vice versa.Can be dissociated with the form of hydrate or the solvate containing solvents used for crystallization Form or the compound of salt form.Formula (I) or the compound of (II) can be reclaimed in a usual manner from reactant mixture and incite somebody to action Its purification.In a usual manner, such as can pass through fractional crystallization or asymmetric synthesis by the such as rotation of corresponding Asymmetrical substitute Photoactive parent material obtains isomer such as stereoisomer.

The compound of formula (I) or (II) can such as use the reaction described in described below and embodiment and technology system Standby.These reactions and can be suitable in the solvent of conversion to be achieved carrying out being suitable for agents useful for same and material.Organic conjunction Skilled artisan understands that of one-tenth field, the functional group being present on molecule should be consistent with the conversion proposed.Sometimes this Need to judge with the change order of synthesis step or select a kind of to be better than alternative ad hoc approach flow process to obtain required basis The compound of invention.

Various substituent groups on the following synthetic intermediate shown in reaction process and end-product are people in the art Member thinks can be presented in fully being modified by suitable protection group in the case of needs, or can be can pass through subsequently Method well-known to those skilled in the art is modified to the precursor forms of its final form and exists.Substituent group can also be in synthesis The different phase of order is added or is added after synthesis order completes.In many cases, conventional functional group manipulations Can be used for one intermediate changes into another kind of intermediate, or the compound of a kind of formula (I) or (II) is changed into another kind Formula (I) or the compound of (II).The example of this generic operation has: ester or ketone are changed into alcohol；Ester is changed into ketone；Ester, acid and acyl The mutual conversion of amine；The alkylation of alkohol and amine, acylated and sulfonylation etc..Popular response such as alkylation, acylated, halogen can also be used Change or substituent group is added in oxidation.This generic operation is well-known in the art, and many reference workss summarize the journey of this generic operation Sequence and method.Give other organic synthesis converted conventional in many functional group manipulations and organic synthesis field The example of main literature and some reference workss of reference are March ' s Organic Chemistry, the 5th edition, Wiley and Chichester edits (2001)；Comprehensive Organic Transformations, Larock edits, VCH (1989)；Comprehensive Organic Functional Group Transformations, Katritzky et al. (series editor), Pergamon (1995)；With Comprehensive Organic Synthesis, Trost and Fleming (it is Row editor), Pergamon (1991).Also it is recognized that arbitrarily another kind of main examining in route of synthesis in plan this area Worry is the protection group of the reactive functionality that careful selection is present in compound of the present invention for protection.Can select same Multiple protection groups in a part, in order to these protection groups each can be in the feelings of other protection group in not removing same a part Being removed under condition and same reactions steps maybe can be used to remove several protection groups, this depends on required result.For holding of undergoing training It is Greene and Wuts that industry personnel describe authority's report of many alternative selections, Protective Groups in Organic Synthesis,Wiley and Sons(1999)。

Pharmacological activity

In terms of regulation CFTR activity, the compound of the formula (I) of free form or pharmaceutical acceptable salt is (hereinafter also referred to as " activating agent of the present invention ") can be used for treating the disease having response to regulation CFTR activity, particularly have benefited from mucosal hydration Disease, such as cystic fibrosis.

Included by the disease of regulation CFTR activity mediation and regulate the disease relevant by the fluid volume of epithelial membrane.Example As, the volume of Airway surface liquid is mucociliary clearance and the critical regulatory factor maintaining lung health.Regulation CFTR activity will Promote the fluid cumulative on the mucosa side of airway epithelia, thus promote mucociliary clearance and prevent mucus and expectorant in respiratory tissue Accumulation in (including lung airway).This kind of disease includes respiratory system disease, such as cystic fibrosis, Primary Ciliary motion barrier Hinder, chronic bronchitis, chronic obstructive pulmonary disease (COPD), asthma, respiratory tract infection (acute and chronic；Viral With bacteroidal) and pulmonary carcinoma.Also include regulating phase with by the abnormal fluid of epithelium by the disease of regulation CFTR activity mediation Disease beyond the respiratory system disease of the abnormal physiological of protectiveness surface liquid that close, that may relate on its surface, example Such as Sjogren syndrome, xerostomia (dry mouth) or keratoconjunctivitis sicca (xerophthalmia).Additionally, the CFTR in regulation kidney lives Performance is used for promoting diuresis and therefore inducing hypotensive activity.

The treatment of the present invention can be for symptom or preventative.

Asthma includes endogenous (anallergic) asthma and exogenous (allergia) asthma, mild asthma, moderate asthma, weight The asthma that degree asthma, bronchitic asthma, the asthma of exercise induced, occupational asthma and antibacterial induce after infecting.Asthma Treatment be also understood as including to e.g., less than 4 years old or 5 years old, demonstrate wheezing symptoms and be diagnosed as maybe can being diagnosed as " wheezy infants " (patient class that a kind of established Major medical is paid close attention to the most often is accredited as the initial stage or roars in early days Asthma) individual treatment.(for convenience, this specific asthma is referred to as that " infant is stridulated syndrome (wheezy infant syndrome)”。)

Preventative effect for the treatment of asthma is by by reducing paresthesia epilepsy, such as acute asthma or the outbreak of bronchoconstriction Frequency and the order of severity, improve pulmonary function or improve airway hyperreactivity and confirm.It can also be right to other by reducing Disease treatment (be used for the most when a symptom occurs or be intended to the treatment for limiting or stop paresthesia epilepsy), such as antiinflammatory are (such as cortex Steroid) or bronchiectasic demand prove.The prevention benefit of asthma is probably spy for easily there is the patient of " morning dipping " The most obvious.The Asthma Syndrome that " morning dipping " is well recognized as, is common in the asthma of significant proportion, it is characterised in that in such as morning Time (the suiting the medicine to the illness property treating asthma time farther out i.e. generally used before any) asthma attack of about 4-6 point.

Chronic obstructive pulmonary disease includes chronic bronchitis or associated dyspnea, emphysema and other medicines Treatment, the airway hyperreactivity that particularly other Drug therapy sucked is caused deteriorate.Present disclosure additionally applies for treating any Type or the bronchitis of origin, including such as acute bronchitis, arachidic bronchitis, Catarrhal bronchus Inflammation, croupous bronchitis, chronic or phthinoid bronchitis.

Xerophthalmia is characterised by that tear produces and reduces and tear film lipid, protein and mucin abnormal distribution.Xerophthalmia is deposited In many reasons, some of which includes age, laser eye surgery, arthritis, medication, chemicals/thermal burn, allergia and disease Sick such as cystic fibrosis and Sjogren syndrome.Increase and can promote that fluid is from corneal epithelial cell by the anion secretion of CFTR With circumocular eccrine transport to increase cornea aquation.This contributes to alleviating the symptom relevant to xerophthalmia.

Sjogren syndrome is a kind of autoimmune disease, wherein the gland producing moisture content in immune system attack body Body, including eye, mouth, skin, respiratory tissue, liver, vagina and intestinal.Symptom includes xerophthalmia, dry mouth and vagina drying and lung disease Sick.This disease is also relevant to rheumatoid arthritis, systemic lupus erythematosus (sle), systemic sclerosis and polymyositis/dermatomyositis.Recognize This disease is caused for protein import defect, limited to its therapeutic choice.CFTR active regulator can make to be encroached on by this disease Various organ aquations and contribute to alleviating relevant symptom.

CFTR active regulator can be tested by measuring the motion of chloride ion in applicable algoscopy based on cell Fitness as the treatment of the disease having benefited from mucosal hydration.Such as, endogenous expression CFTR or be modified to process LAN The epithelial cell that is unicellular or that converge of CFTR can use electrophysiological technique or the research of ion stream to evaluate channel function.See Hirsh et al., J Pharm Exp Ther (2004)；In Moody et al., Am J Physiol Cell Physiol (2005) Described method.

CFTR active regulator including formula (I) compound is alternatively arranged as Co-therapeutic agents and other medicines such as antiinflammatory Medicine, bronchodilator, antihistaminic or cough medicine are applied in combination, particularly at cystic fibrosis or obstructive or inflammatory airway In the treatment of disease (such as those described above), such as this kind of medicine therapeutic activity reinforcing agent or as subtracting The required dosage of few this kind of medicine or the means of potential side effect.

The compound of formula (I) or (II) can be mixed in fixed drug compositions with other medicines or can be by it Before other medicines are used, use the most respectively.

Accordingly, as another aspect, the present invention includes CFTR active regulator and osmotically active agent (osmotic Agent) (saline of high, dextran, mannitol, xylitol), ENaC blocker, anti-inflammatory agent, bronchodilator, anti-group Amine medicine, cough medicine, antibiotic and/or the combination of deoxyribonuclease medicine, wherein CFTR active regulator and other medicine Thing can be in identical or different pharmaceutical composition.

The antibiotic being suitable for includes macrolide antibiotic, such as tobramycin (TOBI^TM)。

The deoxyribonuclease medicine being suitable for includes Dornase Alfa (Pulmozyme^TM) a kind of highly purified heavy The solution of group people's deoxyribonuclease I (rhDNase), its selective splitting DNA.Dornase Alfa is used for treating capsule fiber Change.

CFTR active regulator with other useful combination of anti-inflammatory agent is and chemokine receptor anagonists, such as CCR- 1, CCR-2, CCR-3, CCR-4, CCR-5, CCR-6, CCR-7, CCR-8, CCR-9 and CCR10, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, particularly CCR-5 antagonist, as Schering-Plough antagonist SC-351125, SCH-55700 and The antagonist of SCH-D, Takeda such as N-[[4-[[[6,7-dihydro-2-(4-methylphenyl)-5H-benzo-cycloheptene-8-base] Carbonyl] amino] phenyl]-methyl] tetrahydrochysene-N, N-dimethyl-2H-pyrans-4-ammonium chloride (TAK-770) and USP6,166, (particularly right is wanted for 037 (particularly claim 18 and 19), WO00/66558 (particularly claim 8), WO00/66559 Ask 9), the combination of the CCR-5 antagonist described in WO04/018425 and WO04/026873.

The anti-inflammatory agent being suitable for includes: steroid, particularly glucocorticoid, such as budesonide, beclometasone, Fluticasone propionate, ciclesonide or momestasone furoate, or the steroid described in following documents: WO02/88167, WO02/ 12266, WO02/100879, WO02/00679 (especially embodiment 3,11,14,17,19,26,34,37,39,51,60,67, 72, those of 73,90,99 and 101), WO03/35668, WO03/48181, WO03/62259, WO03/64445, WO03/ 72592, WO04/39827 and WO04/66920；On-steroidal glucocorticoid receptor agonist, such as described in following documents Those: DE10261874, WO00/00531, WO02/10143, WO03/82280, WO03/82787, WO03/86294, WO03/104195, WO03/101932, WO04/05229, WO04/18429, WO04/19935 and WO04/26248；LTD4 antagonism Agent, such as montelukast and zafirlukast；PDE4 inhibitor, such as cilomilast (GlaxoSmithKline), sieve Fluorine department special (Byk Gulden), V-11294A (Napp), BAY19-8004 (Bayer), SCH-351591 (Schering- Plough), arofylline (Almirall Prodesfarma), PD189659/PD168787 (Parke-Davis), AWD-12- 281(Asta Medica)、CDC-801(Celgene)、SelCID(TM)CC-10004(Celgene)、VM554/UM565 (Vernalis), T-440 (Tanabe), KW-4490 (Kyowa Hakko Kogyo) and WO92/19594, WO93/19749, WO93/19750、WO93/19751、WO98/18796、WO99/16766、WO01/13953、WO03/104204、WO03/ 104205、WO03/39544、WO04/000814、WO04/000839、WO04/005258、WO04/018450、WO04/ 018451、WO04/018457、WO04/018465、WO04/018431、WO04/018449、WO04/018450、WO04/ 018451, WO04/018457, WO04/018465, WO04/019944, WO04/019945, WO04/045607 and WO04/ Those disclosed in 037805；Those described in adenosine A 2B receptor antagonist, such as WO02/42298；With β-2 epinephrine Receptor stimulating agent, such as albuterol (salbutamol), orciprenaline, terbutaline, salmaterol, fenoterol, procaterol Especially the formula (I) in formoterol, carmoterol or its officinal salt, and WO0075114 compound (free form or Salt form or solvate form thereof), by reference the document is integrated with herein, the preferably compound of embodiment, especially It is following formula

Corresponding to QAB-149 or its officinal salt, and compound (free form or the salt of the formula (I) of WO04/16601 Form or solvate form thereof), also the compound in documents below: EP1440966, JP05025045, WO93/18007, WO99/64035、USP2002/0055651、WO01/42193、WO01/83462、WO02/66422、WO02/70490、WO02/ 76933、WO03/24439、WO03/42160、WO03/42164、WO03/72539、WO03/91204、WO03/99764、WO04/ 16578、WO04/22547、WO04/32921、WO04/33412、WO04/37768、WO04/37773、WO04/37807、WO04/ 39762, WO04/39766, WO04/45618, WO04/46083, WO04/80964, WO04/108765 and WO04/108676.

The bronchodilator being suitable for includes anticholinergic or antimuscarinic drug, particularly ipratropium bromide, oxygen torr bromine Ammonium, tiotropium salt and CHF4226 (Chiesi) and glycopyrronium bromide, also those described in documents below: EP424021, USP3,714,357、USP5,171,744、WO01/04118、WO02/00652、WO02/51841、WO02/53564、WO03/ 00840, WO03/33495, WO03/53966, WO03/87094, WO04/018422 and WO04/05285.

The dual anti-inflammatory and the bronchodilator that are suitable for include that dual beta-2-adrenoceptor agonist/muscarine is short of money Anti-agent, such as those disclosed in USP2004/0167167, WO04/74246 and WO04/74812.

Be suitable for antihistamine drug include cetirizine hydrochloride, acetaminophen, clemastine fumarate, promethazine, Loratadine, Desloratadine, diphenhydramine and fexofenadine hydrochloride, acrivastine (activastine), astemizole, NitrogenSTING, ebastine, epinastine, mizolastine and tefenadine (tefenadine), and JP2004107299, Those disclosed in WO03/099807 and WO04/026841.

As other side, the present invention comprises the combination of CFTR active regulator and CFTR corrigent, wherein CFTR activity Regulator and CFTR corrigent can be in identical or different pharmaceutical compositions.The CFTR corrigent being suitable for includes VX-809

With

VX-661

As described above, as another aspect, present invention also offers treatment has response to regulation CFTR activity Disease, such as to the regulation method by the relevant disease of the fluid volume of epithelial membrane, particularly obstructive airway diseases, the party Method include to needs its individuality, particularly individual human use formula (I) or the change of (II) of free form or pharmaceutical acceptable salt Compound.

In yet another aspect, present invention provide for preparing the formula (I) of the free form of medicine or pharmaceutical acceptable salt Or the compound of (II), described medicine has the disease of response, particularly obstructive airway disease for treatment to regulation CFTR activity Disease, such as cystic fibrosis and COPD.

The activating agent of the present invention can by any suitable by way of using, the most Orally administered, such as with tablet or glue The form of capsule is Orally administered；Parenteral administration, such as intravenous are used；Used by suction, such as at obstructive airway diseases Treatment in；Intranasal administration, such as in the treatment of allergic rhinitis；It is locally applied to skin；Or rectal administration.At another Aspect, present invention also offers pharmaceutical composition, and it comprises formula (I) compound of free form or pharmaceutical acceptable salt, optionally Comprise pharmaceutically useful diluent or carrier.Described compositions can contain Co-therapeutic agents, anti-inflammatory agent the most mentioned above, a gas Enlargement of pipe medicine, antihistaminic or cough medicine.This based composition can use known conventional diluent or tax in formulation art Prepared by shape agent and technology.Therefore, peroral dosage form can include tablet and capsule.Formulations for topical administration can use breast The form of unguentum, ointment, gel or transdermal delivery system such as patch.Composition for inhalation can include aerosol or its Its atomisable formulation or dry powder formulations.

When compositions includes aerosol formulation, it preferably comprises such as hydrogen-fluoro-alkane (HFA) propellant, such as HFA134a or HFA227 or their mixture, and one or more cosolvent known in the art, such as second can be contained Alcohol (at most 20% weight), and/or one or more surfactants, such as oleic acid or Sorbitan Trioleate, and/or a kind of or Multiple filler, such as lactose.When compositions includes dry powder formulations, it preferably comprises such as has at most 10 micron grain sizes Formula (I) or the compound of (II), optionally the diluent or carrier such as lactose containing the distribution of required particle diameter, and contribute to preventing product The compound that performance deteriorates because of dampness, such as magnesium stearate.When compositions includes atomization preparation, it preferably comprises and is such as dissolved in Or the formula (I) that is suspended in vehicle or the compound of (II), described vehicle includes water, cosolvent such as ethanol or the third two Alcohol and stabilizer, stabilizer can be surfactant.

Other aspects of the present invention include:

(a) inhalable form, such as aerosol or other nebulizable composition or inhalable particles such as Micronised form Formula (I) or the compound of (II)；

B () comprises the formula (I) of inhalable form or the sucked medicament of the compound of (II)；

C () comprises compound and the drug products of suction apparatus of the formula (I) of inhalable form；With

(d) compound containing the Formulas I of inhalable form or II can suction apparatus.

Certainly concrete disease basis such as treated for the dosage of the compound of the formula (I) or (II) of implementing the present invention Disease, required effect and the difference of method of application and change.It is said that in general, exist for sucking the daily dose being suitable for used 0.005-10mg rank, and for the Orally administered daily dose being suitable in 0.05-100mg rank.

Medicinal usage and algoscopy

Formula (I) or the compound of (II) and officinal salt (hereinafter also referred to " activating agent of the present invention ") thereof can be used as medicine Thing.Especially, described compound is applicable CFTR active regulator and can test in following assay.

Transmembrane potential algoscopy

By measuring transmembrane potential, CFTR activity can be carried out quantitatively.The method of the transmembrane potential in measurement biosystem Can make in many ways, including the transmembrane potential algoscopy of electrophysiology and optically-based fluorescence.

Optical membrane potential assay uses electronegative potential measurement dyestuff, such as FLIPR transmembrane potential dyestuff (FMP) (see Baxter DF, Kirk M, Garcia AF, Raimondi A, Holmqvist MH, Flint KK, Bojanic D, New for Distefano PS, Curtis R, Xie Y. ' transmembrane potential sensitive fluorescent dye improve for ion channel based on carefully Algoscopy (the A novel membrane potential-sensitive fluorescent dye improves of born of the same parents Cell-based assays for ion channels). ' J Biomol Screen.2002 February；7 (1): 79-85), it Quencher is combined when born of the same parents are outer.After cell depolarization, electronegative dyestuff reassigns to intracellular compartment, thus non-from film Infiltration quencher discharges, causes fluorescence to increase.This fluorescence change with may changing because of transmembrane potential that CFTR activity causes Become direct ratio.Can be micro-in 96 or 384-holes by the fluorescence detector such as FLIPR (Fluorometric Imaging Plate reader) of proper distrabution Amount titer plate is monitored the change of fluorescence in real time.

Cell is cultivated:

Transmembrane potential experiment is carried out with Chinese hamster ovary (CHO) cell of stable expression Δ F508-CFTR passage.By cell In 37 DEG C, at 5%v/v CO₂In, maintain in MEM (MEM) under 100% humidity, this culture media supplemented 8%v/v hyclone, 100 μ g/ml methotrexate and 100U/ml penicillin/streptomycin.Cell is made to be grown in 225cm²Group Knit in culture bottle.In order to carry out transmembrane potential mensuration, by cell with 40,000 cells/well is seeded in 96 orifice plates so that it is adhere to, so After maintain 26 DEG C reach 48h so that passage insert.

Reinforcing agent algoscopy:

Transmembrane potential Screening test method make use of the low chlorine ion (～5mM) containing the outer solution of born of the same parents and two-cocoon feeding to add scheme.The Once adding is the buffer with or without test compound, and adding Forskolin (1-20 μM) program after 5 minutes has It is beneficial to flow out in response to the maximum chlorine of Δ F508-CFTR activation.The chloride ion of Δ F508-CFTR mediation flows out and causes film to go to pole Change, optionally through FMP dyestuff, it is monitored.

Solution:

The outer solution (mM) of low chlorine born of the same parents: 120 gluconic acid Na, 1.2CaCl₂,3.3KH₂PO₄,0.8K₂HPO₄,1.2MgCl₂, 10.0D-glucose, 20.0HEPES, it is adjusted to pH7.4 with NaOH

FMP dyestuff: preparing the outer solution of above-mentioned low chlorine born of the same parents according to the explanation of manufacturer, 10x final concentration, with 1mL aliquot It is stored in-20 DEG C.

IonWorks Quattro algoscopy:

CFTR activity can be carried out quantitatively in electrophysiology mode to use the patch clamp technique of full cell configuration (Hamill et al. Pflugers Acrhive1981).This algoscopy is directly measured relevant to by the chlorine stream of CFTR passage Electric current, maintains simultaneously or adjusts transmembrane voltage.This algoscopy can use single glass micropipettor or parallel planar arrays Measure the CFTR activity from natural or reconstitution cell system.The instrument such as IonWorks of suitably outfit can be used It is fixed that the electric current using parallel planar arrays to measure is carried out by Quattro (Molecular Devices) or Qpatch (Sophion) Amount.Quattro system can be measured from unicellular/register hole (HT structure) or the group/hole from 64 cells The CFTR electric current of (Population Patch Clamp PPC) (Finkel A, Wittel A, Yang N, Handran S, Hughes J, Costantin J. ' group's patch-clamp improves data consistency and success rate (Population in measuring ion stream patch clamp improves data consistency and success rates in the measurement of Ionic currents). ' J Biomol Screen.2006 August；11 (5): 488-96).

Cell is cultivated:

IonWorks Quattro is carried out with Chinese hamster ovary (CHO) cell of stable expression Δ F508-CFTR passage Experiment.By cell in 37 DEG C, at 5%v/v CO₂In, maintain under 100% humidity and supplement 10% (v/v) FCS, 100U/ ML penicillin/streptomycin, 1% (v/v) NEAA, 1mg/ml Zeocin and 500ug/ml HYG D-MEM in.In order to carry out Experiment, makes cell be grown in 225cm²To close to converging in tissue culture flasks, then cultivate 48-72h so that passage in 26 DEG C Insert.From bottle, take out cell, be resuspended in for carrying out immediately in the Recording out cell solution tested or being resuspended in benefit Fill in the growth medium of 10%v/v DMSO and be refrigerated to-80 DEG C in case use later with 1-2mL aliquot.

Reinforcing agent algoscopy:

Cell is placed in Quattro system with the density of 1.5-3 million/mL, joins in plane patch array, close Envelope so that it is set up 5-10 minute.After evaluating and sealing resistance (generally > 50M Ω), by boring a hole with 100 μ g/mL amphotericin Bs Obtain full cell pathway.By scanning survey base current before apply the compound that voltage obtains from-100 to+100mV.So After test compound by buffer or with the born of the same parents supplementing 20 μMs of Forskolins outer solution dilution join plane patch array 384 holes each hole in.After incubation step (5-20 minute), again measure by applying voltage from-100 to+100mV Compound after current.Difference between current between scanning before compound and after compound defines the effect that CFTR strengthens.

Solution:

The outer solution (ECS) of born of the same parents: 145mM NaCl, 4mM CsCl, 5mM D-Glucose, 10mM TES, 1mM CaCl₂,1mM MgCl₂,pH7.4NaOH

Intracellular buffer (ICS): 113mM L-Aspartic acid, 113mM CsOH, 27mM CsCl, 1mM NaCl, 1mM MgCl₂, 1mM EGTA, 10mM TES. CsOH is adjusted to pH7.2.Aseptic filtration is carried out before using.

Ion transport algoscopy:

The another kind of method measuring CFTR function is Ussings room short circuit current measurement.Make transformation or natural epithelium Cell is grown to the monolayer that converges on semi-transparent filter membrane and is clipped between two methyl methacrylate (perspex) blocks.Can With the flow by measurement electric current, the chloride ion flow flowing to opposite side via the side from epithelium of CFTR is carried out quantitatively, with Time transepithelial current potential is maintained 0mV.This is that the electrode based on agar using KCl to fill is to vise cell monolayer and to measure Current flow and realize.

Cell is cultivated:

The FRT cell of stable expression Δ F508-CFTR is being supplemented 32mM NaHCO as growth medium₃、 10%v/v hyclone, 2mM L-glutaminate, 100U/mL penicillin, 100 μ g/mL streptomycins and 30 μ g/mL HYGs Cultivate in the F-12 culture medium that Coon improves.In order to carry out the experiment of Ussing room, cell is made to exist as the epithelial growth of polarization Snapwell permeability is supported, on insert (500000 cell/inserts in growth medium), to cultivate 7-9 days.Often 48 hours and test first 24 hours and fill, carrying out Ussing room, the F-12 grown cultures that fresh Coon improves to insert Base.In order to increase Δ F508CFTR protein expression on cell surface, by plate in 27 DEG C of incubation 48h, then carry out Ussing room Experiment.

Reinforcing agent algoscopy:

The Fischer of the stable expression people Δ F508-CFTR of use monolayer cultures form on permeable holder Rat thyroid (FRT) epithelial cell.In Ussing room use short circuit current technology, force from substrate side to top Cl^-Cl is measured under gradient^-Electric current.In order to measure stable Cl^-Electric current, cultivates 48h so that Δ by FRT cell at 27 DEG C F508CFTR inserts plasma membrane.The research of Ussing room is carried out at 27 DEG C equally.Under these conditions, it is possible to use titer and effect are eventually Point quantitatively accumulates the interpolation test compound effect to Δ F508CFTR electric current.By compound after adding 10 μMs of Forskolins Join top and substrate side.The effect of compound is compared with known reinforcing agent such as Gensitein.

Solution:

Substrate side ringer's solution (mM): 126NaCl, 24NaHCO₃,0.38KH₂PO₄,2.13K₂HPO₄,1MgSO₄,1CaCl₂ With 10 glucoses.

Top ringer's solution (mM): 140 gluconic acid Na, 1MgSO₄,2CaCl₂, 1HCl, 10 glucoses and 24NaHCO₃。

Said determination method test compound can also be used to stimulate Δ F508CFTR to insert the ability of cell membrane.For these Algoscopy, scheme is identical, except for the difference that cell not under low temperature (26 or 27 DEG C) cultivate, but before the assay with testization Compound incubation 12-24h together.

Generally, the compound of Examples below has the EC less than 10 μMs in above-mentioned DATA REASONING₅₀Value.Table 1 Provide list and the EC thereof of representative compound₅₀Value.

Table 1.

Embodiment is numbered	EC₅₀μM	Embodiment is numbered	EC₅₀
				1	0.0023829	10	0.0014
2	0.003408	10.1	0.013
				3	0.00355	10.2	0.005227
4	0.15	10.3	0.0743833
				5	0.0866667	10.4	0.0313
6	0.1845	10.5	0.127
				7	0.0041	10.6	0.0146625
8	0.0021	11	0.078
				9	0.0845	12	0.010

The present invention is illustrated by the following example.

Embodiment

General conditions:

The LCMS system using electron spray ionisation is run mass spectrum.These are Agilent1100HPLC/Micromass Platform Mass Spectrometer combines or with SQD mass spectrometric Waters Acquity UPLC.[M+H]⁺Refer to List-isotopic molecule amount.

The open entrance Bruker AVANCE400NMR spectrophotometer using ICON-NMR runs NMR spectra.? 298K measure spectrum and use solvent peak as reference.

The following example is used for illustrating the present invention, but should not be understood as limitation of the present invention.Temperature is to take the photograph Family name's degree is given.Without mentioning other condition, the most all of evaporation is the most under reduced pressure carried out, preferably about 15 Carry out under mm Hg-100 mm Hg (=20-133 mbar).The structure of end-product, intermediate and parent material is to pass through Standard method of analysis, such as microanalysis and spectrophotometric feature, such as MS, IR, NMR confirm.Abbreviation used is ability Those abbreviations conventional in territory.If it is not defined, then term has its implication generally accepted.

Abbreviation:

App is apparent

BEMP 2-tertbutylimido-2-diethylamino-1,3-dimethyl is complete

Hydrogenation-1,3,2-diaza phospha benzene

Boc tert-butoxycarbonyl

Br broad peak

D is bimodal

Dd double doublet

DCM dichloromethane

DIPEA diisopropylethylamine

DMF N,N-dimethylformamide

DMSO dimethyl sulfoxide

EtOAc ethyl acetate

H hour

HATU 2-(7-azepine-1H-benzotriazole-1-base)-1,1,3,3-tetramethyl

UreaHexafluorophosphate

HPLC high pressure lipuid chromatography (HPLC)

Int. intermediate

LC-MS liquid chromatography and mass spectrography

MeOH methanol

MS mass spectrography

M multiplet

Min minute

Ml milliliter

M/z mass-to-charge ratio

NMR nuclear magnetic resonance, NMR

Ppm 1/1000000th

PS polymer is supported

PEAX PE-anion exchange is (such as from Biotage's

PE-AX post)

RT room temperature

Rt retention time

S is unimodal

SCX-2 strong cation exchange is (such as from Biotage's

SCX-2 post)

T triplet

TEA triethylamine

TFA trifluoroacetic acid

THF oxolane

With reference to the following examples, method described herein or other method known in the art is used to synthesize preferred reality Execute the compound of scheme.

In a suitable manner, it is possible to use routine techniques such as precipitates, filter, crystallize, evaporate, distill and chromatography divides From various parent materials, intermediate and compound with purification preferred embodiment.Unless otherwise stated, all initial materials Material is all obtained from commercial supplier and the most i.e. uses.Can be prepared by compound by the operation of known one-tenth salt Salt.

It should be appreciated that the organic compound of preferred embodiment can show tautomerism.Due to this explanation Chemical constitution in book is only capable of one of tautomeric form of expressing possibility, it is, therefore, appreciated that preferred embodiment bag Include any tautomeric form of shown structure.

If not stated otherwise, then analytical type HPLC condition is as follows:

Method 10minLC_v002

Post Waters BEH C1850x2.1mm, 1.7 μm

Column temperature 50 DEG C

Eluant A:H₂O, B: methanol, all contains 0.1%TFA

Flow velocity 0.8ml/min

Gradient 0.20min5%B；In 5%-95%B, 7.80min, 1.00min95%B

Method 10minLC_v003

Post Waters BEH C1850x2.1mm, 1.7 μm

Column temperature 50 DEG C

Eluant A:H₂O, B: acetonitrile, all contains 0.1%TFA

Flow velocity 0.8mL/min

Gradient 0.20min5%B；In 5%-95%B, 7.80min, 1.00min95%B

Method 2minLC_v002

Post Waters BEH C1850x2.1mm, 1.7 μm

Column temperature 50 DEG C

Eluant A:H₂O, B: methanol, all contains 0.1%TFA

Flow velocity 0.8ml/min

Gradient 0.20min5%B；In 5%-95%B, 1.30min, 0.25min95%B

Method 2minLC_v003

Post Waters BEH C1850x2.1mm, 1.7 μm

Column temperature 50 DEG C

Eluant A:H₂O, B: acetonitrile, all contains 0.1%TFA

Flow velocity 0.8ml/min

Gradient 0.20min5%B；In 5%-95%B, 1.30min, 0.25min95%B

Method HighpH_v003

Post Waters BEH C1850x2.1mm, 1.7 μm

Column temperature 50 DEG C

Eluant A:H₂O, B: acetonitrile, all contains 0.1%NH₃OH

Flow velocity 0.8mL/min

Gradient 0.25min5%B；In 5%-95%B, 1.00min, 0.25min95%B

Embodiments of the invention compound includes:

The preparation of finalization compound

Embodiment 1.0

2-(5-(3-amino-6-methoxyl group-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1, 1-trifluoro propan-2-ol

Step 1:2-(2-(the fluoro-2-methylpropionyl of 2-(benzyloxy)-3,3,3-three) hydrazine carbonyl)-6-methoxyl group-5-(three Methyl fluoride) pyridin-3-yl carbamate

To the fluoro-2 Methylpropionic acid of 2-(benzyloxy)-3,3,3-three (intermediate D) (0.709g, 2.85mmol) at DMF (15ml) solution adds HATU (1.303g, 3.43mmol), is subsequently added DIPEA (0.598ml, 3.43mmol).30 points Zhong Hou, adds 2-(hydrazine carbonyl)-6-methoxyl group-5-(trifluoromethyl) pyridin-3-yl carbamate (intermediate C) (1g, 2.85mmol), and reactant mixture is stirred under RT 3h.This mixture is poured into water, and produces with EtOAc extraction Thing.By organic moiety 1MHCl, saline washing, it is dried (MgSO₄) and be concentrated in vacuo.Through silica gel chromatography 0%-50%EtOAc Eluant solution purification in isohexane, obtains title compound:

Step 2:2-(5-(2-(benzyloxy)-1,1,1-trifluoropropyl-2-base)-1,3,4-diazole-2-base)-6-methoxy Base-5-(trifluoromethyl) pyridin-3-yl carbamate

2-(2-(the fluoro-2-methylpropionyl of 2-(benzyloxy)-3,3,3-three) hydrazine carbonyl)-6-methoxyl group-5-by stirring (trifluoromethyl) pyridin-3-yl carbamate (step 1) (1g, 1.723mmol) mixing in anhydrous THF (20ml) Suspension burgess reagent (3 equivalent) processes, and at N₂Under be heated at reflux 1 hour 45 minutes.RM is concentrated in vacuo to about 5ml Volume, and dilute with EtOAc (150ml).By this mixture 2M NaOH, 0.5M HCl, water, saline washing, it is dried (MgSO₄) and be concentrated in vacuo.Crude product MeOH is ground, obtains title compound.LC-MS Rt=1.59min [M+H]+ 563.5 method High pH_v003.

Step 3:2-(5-(2-(benzyloxy)-1,1,1-trifluoropropyl-2-base)-1,3,4-diazole-2-base)-6-methoxy Base-5-(trifluoromethyl) pyridine-3-amine

2-(5-(2-(benzyloxy)-1,1,1-trifluoropropyl-2-base)-1,3,4-diazole-2-base)-6-first in stirring Epoxide-5-(trifluoromethyl) pyridin-3-yl carbamate (707mg, 1.257mmol) is at the solution of DCM (10ml) Middle addition TFA (3ml), and continue to stir 45 minutes.Gained mixture 2M NaOH is washed, and has with phase separator separation Machine phase.Solvent removed in vacuo, obtains title compound, for tfa salt；LC-MS Rt=1.42min [M+H]+463.2 method HighpH_v003。

Step 4:2-(5-(3-amino-6-methoxyl group-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2- Base)-1,1,1-trifluoro propan-2-ol

2-(5-(2-(benzyloxy)-1,1,1-trifluoropropyl-2-base)-1,3,4-diazole-2-base)-6-first in stirring Epoxide-5-(trifluoromethyl) pyridine-3-amine trifluoroacetate (step 3) (450mg, 0.973mmol) is molten EtOH's (25ml) Liquid adds palladium dydroxide/carbon (45.1mg, 0.321mmol), is subsequently added ammonium formate (614mg, 9.73mmol).By this mixing Thing is heated at reflux 1h, and with after warpFilter, with EtOH and be subsequently washed with water.Ethanol removed in vacuo, and by gained Aqueous phase EtOAc extracts.By the organic extract use water merged, saline washing, it is dried (MgSO4) and is concentrated in vacuo, being marked Topic product；1H NMR(400MHz,DMSO-d6)δ7.85(1H,s),7.74(1H,s),6.74(2H,s),3.94(3H,s), 1.86 (3H, s) .LC-MS Rt=1.17min [M+H]+373.1 method: 2minLC_v003.

Embodiment 2 and 3

Use supercritical fluid chromatography by 2-(5-(3-amino-6-methoxyl group-5-(trifluoromethyl) pyridine-2-base)-1, 3,4-diazole-2-bases)-1,1,1-trifluoro propan-2-ol (embodiment 1) chiral separation, obtain enantiomer 1 and 2:

Condition:

Flowing phase: 10%MeOH+0.1%DEA/90%CO2

Post: Chiralpak AD-H, 250x10mm id, 5 μm

Detection: UV@220nm

Flow velocity: 10ml/min

Sample concentration: 63mg/ml (250mg+4ml EtOH).

Embodiment 2:First eluting peak.Enantiomer 1:(R)-2-(5-(3-amino-6-methoxyl group-5-(fluoroform Base) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoro propan-2-ol

1H NMR(400MHz,DMSO-d6)δ7.86(1H,s),7.75(1H,s),6.74(2H,s),3.94(3H,s), 1.86 (3H, s) .LC-MS Rt=4.04min [M+H]+373.4 (method 10min_v003).The spatial chemistry warp of this compound 3D X-ray crystallographic data confirms.

Embodiment 3:Second eluting peak.Enantiomer: (S)-2-(5-(3-amino-6-methoxyl group-5-(fluoroform Base) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoro propan-2-ol

1H NMR(400MHz,DMSO-d6)δ7.86(1H,s),7.75(1H,s),6.74(2H,s),3.94(3H,s), 1.86 (3H, s) .LC-MS Rt=4.04min [M+H]+373.4 methods 10min_v003.

Embodiment 4

3-(5-benzyl-1,3,4-diazole-2-base)-5-bromo-6-(trifluoromethyl) pyrazine-2-amine

Step 1: 3-amino-6-bromo-5-(trifluoromethyl) pyrazine-2-formylhydrazine

Exist to 3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-Ethyl formate (intermediate F1) (1.00g, 3.18mmol) The solution of anhydrous EtOH (25ml) adds monohydrate hydrazine (309ul, 6.37mmol), and by under the backflow of this mixture (about 90 DEG C) stirring.After 3 hours 45 minutes, reactant mixture is cooled down in ice bath, and is added to the water.By gained yellow mercury oxide vacuum Filter, and be dried overnight in a vacuum furnace, obtain title compound, for yellow solid.¹H NMR(400MHz,DMSO-d6)δ 10.10(1H,s),8.00(2H,s),4.65(2H,s)。

Step 2:The bromo-N'-of 3-amino-6-(2-phenyl acetyl)-5-(trifluoromethyl) pyrazine-2-formylhydrazine

In the solution of NMP (6ml), 3-amino-6-is added bromo-to stirred phenylacetic acid (95.0mg, 0.698mmol) 5-(trifluoromethyl) pyrazine-2-formylhydrazine (step 1) (250mg, 0.837mmol) solution in NMP (6ml), is subsequently added DIPEA(609ul,3.49mmol).By HATU (398mg, the 1.047mmol) batch processing of gained mixture, and stir under RT Mix 90 minutes.Add the mixture in EtOAc (50ml), and wash with 0.1M NaOH (50ml).Water layer is used again EtOAc (50ml) extraction.By organic moiety use water (50ml) merged, saline (50ml) washing, it is dried with magnesium sulfate, and is concentrated in vacuo. By crude on silica gel chromatographic grade 1:1EtOAc/ iso-hexane purification, obtain title compound, for yellow solid.1H NMR (400MHz,DMSO-d6)δ10.65(1H,s),10.29(1H,s),8.02(2H,s),7.30(5H,m),3.52(2H,s)。 19FNMR(400MHz,DMSO-d6)δ-66(s,CF3)。

Step 3:3-(5-benzyl-1,3,4-diazole-2-base)-5-bromo-6-(trifluoromethyl) pyrazine-2-amine

To the 3-bromo-N'-of amino-6-(2-phenyl acetyl)-5-(trifluoromethyl) pyrazine-2-formylhydrazine (step 1) (92mg, 0.220mmol) adds molten in THF (0.5ml) of TsCl (50mg, 1.200mmol) in the solution of THF (2ml) Liquid.Subsequently gained solution is added in PS-BEMP (384mg, 1.100mmol), and by this mixture microwave radiation at 120 DEG C Heat 10 minutes.Reactant mixture is loaded with THF moistening in advance(10g, solid phase extracts PE-AX/SCX-2 post Take) in.By this post THF (40ml) eluting, and organic moiety is concentrated in vacuo.By crude on silica gel chromatographic grade 20% EtOAc/ iso-hexane purification, the most further through preparation HPLC purification, obtains title compound；1H NMR(400MHz, DMSO-d6)δ7.35(5H),4.49(2H,s).LC-MS Rt=5.76min [M+H]+400.1 method 10minLC_v002.

Embodiment 5

(5-(3-amino-6-bromo-5-(trifluoromethyl) pyrazine-2-base)-1,3,4-diazole-2-base) (phenyl) ketone

Prepare title compound according to method similar to Example 4, replace phenylacetic acid (step 2) with phenylglyoxalates； 1H NMR(400MHz,DMSO-d6)δ8.41(2H,d),7.82(1H,t),7.70(2H,t).LC-MS Rt=5.82min [M+ H]+416.1 methods 10minLC_v002.

Embodiment 6

2-(5-(3-amino-5,6-two (trifluoromethyl) pyrazine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoro Propan-2-ol

Step 1:3-amino-N'-(the fluoro-2-methylpropionyl of 2-(benzyloxy)-3,3,3-three)-5,6-two (trifluoromethyl) Pyrazine-2-formylhydrazine

By stirred 3-amino-5,6-two (trifluoromethyl) pyrazine-2-formylhydrazine (intermediate G) (431mg, 1.491mmol) the fluoro-2 Methylpropionic acid of solution 2-(the benzyloxy)-3,3,3-three (intermediate D) in anhydrous NMP (3ml) (370mg, 1.491mmol) processes, and stirs 5 minutes under RT.Gained yellow solution is cooled to 0 DEG C, and uses HATU (567mg, 1.491mmol) processes, and is added dropwise over triethylamine (0.208ml, 1.491mmol) subsequently.By this orange/redness suspension Stir 15 minutes at 0 DEG C, and be heated up to RT subsequently, stir about 2 hours.By this mixture at EtOAc (50ml) and 1M Distribute between NaOH (50ml), shake and separate.Organic moiety saline (25ml) is washed, is dried (MgSO₄) and vacuum dense Contracting, obtains orange.Crude product is eluting through the LC-MS MeCN/ water/0.1%TFA of Mass Spectrometer Method.Will clarification Flow point pour in EtOAc (50ml), and use saturated NaHCO₃Solution (50ml) washs, and is dried (MgSO₄) and be concentrated in vacuo, To title compound, for light yellow crystalline solid.

1H NMR (400MHz, DMSO-d6) δ 10.82 (1H, br s), 10.4 (1H, br s), 8.72 (1H, wide hump), 8.4 (1H, wide humps), 7.5 (2H, d), 7.3-7.42 (3H, m), 4.79 (2H, m), 1.7 (3H, s).(there is trace EtOAc, But it is pure and relevant to the structure supposed).

19F NMR (400MHz, DMSO-d6): peak 1 is at-61ppm, and peak 2 is at-64.5ppm, and peak 3 is at-76ppm

Step 2:3-(5-(2-(benzyloxy)-1,1,1-trifluoropropyl-2-base)-1,3,4-diazole-2-base)-5,6-two (trifluoromethyl) pyrazine-2-amine

By stirred 3-amino-N'-(the fluoro-2-methylpropionyl of 2-(benzyloxy)-3,3,3-three)-5,6-two (trifluoro Methyl) pyrazine-2-formylhydrazine (step 1) (510mg, 0.982mmol) solution triethylamine in DCM (20ml) (0.411ml, 2.95mmol) processes, and stirs under RT.Add toluene sulfochloride (562mg, 2.95mmol), and continue stirring 30 minutes.Reactant mixture is distributed between EtOAc (50ml) and 1M HCl (50ml), shakes and separate.By organic moiety Wash with saline (30ml), be dried (MgSO₄) and be concentrated in vacuo, obtain yellow oil.By crude product through the LC-of Mass Spectrometer Method MS MeCN/ water/0.1%TFA is eluting.This clarification flow point is poured in EtOAc (50ml), and uses saturated NaHCO₃Solution (50ml) washing, is dried (MgSO₄) and be concentrated in vacuo, obtain title compound, for white crystalline solid.

1H NMR (400MHz, DMSO-d6) δ 9.28 (1H, wide hump), 8.1 (1H, wide humps), 7.46 (2H, m), 7.3- 7.4(3H,m),4.72(1H,d),4.52(1H,d),2.1(3H,s).LC-MS Rt=1.40min [M+H]+502.2 method 2minLC_v003。

Step 3:2-(5-(3-amino-5,6-two (trifluoromethyl) pyrazine-2-base)-1,3,4-diazole-2-base)-1,1, 1-trifluoro propan-2-ol

At N₂Under at 3-(5-(2-(benzyloxy)-1,1,1-trifluoropropyl-2-base)-1,3,4-diazole-2-base)-5,6-two (trifluoromethyl) pyrazine-2-amine (step 2) (50mg, 0.100mmol) loads Pd/C in the solution of EtOH (3ml) (1.061mg, 9.97 μm ol), and under RT, it is placed in H2 (0.35bar) direct draught overnight.By reactant mixture warp (filter material) filters, and through ethanol (30ml), with after wash through DCM (10ml).Filter vacuum is concentrated, and by gained crude product warp The LC-MS MeCN/ water/0.1%TFA of UV detection is eluting.This clarification flow point is poured in EtOAc (50ml), and with full And NaHCO₃Solution (50ml) washs, and is dried (MgSO₄) and be concentrated in vacuo, obtain title compound, for orange solid.1H NMR (400MHz, DMSO-d6) δ 9.22 (1H, wide hump), 8.1 (1H, wide humps), 7.92 (1H, s), 1.4 (3H, s).19F NMR (400MHz, DMSO-d6) peak 1 is at-61ppm, and peak 2 is at-64ppm, and peak 3 is at-79.4ppm.

Embodiment 7 and 8

These compounds are i.e.:

According to following HPLC condition by 2-(5-(3-amino-5,6-two (trifluoromethyl) pyrazine-2-base)-1,3,4-two Azoles-2-base)-1,1,1-trifluoro propan-2-ol (embodiment 6) chiral separation prepares (R)-2-(5-(3-amino-5,6-two (three Methyl fluoride) pyrazine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoro propan-2-ol and (S)-2-(5-(3-amino-5,6- Two (trifluoromethyl) pyrazine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoro propan-2-ol:

Post: Chiralcel OJ-H, 20x250mm id, 10 μm

Flowing phase: 4%EtOH/96% heptane+0.1DEA

Flow velocity: 20ml/min

Detection: UV@320nm

Sample injection 250mg solution (3ml) in EtOH

Volume injected: 125 μ l

Embodiment 7 and 8 is enantiomer.Through post isolation identification compound.

Embodiment 7:First eluting peak.2-(5-(3-amino-5,6-two (trifluoromethyl) pyrazine-2-base)-1,3,4- Diazole-2-base) enantiomer 1 of-1,1,1-trifluoro propan-2-ol

1H NMR(400MHz,DMSO-d6)δ9.18-9.3(1H,br s),8.02-8.18(1H,br s),7.92(1H, s),1.89(3H,s).19F NMR (400MHz, DMSO-d6) peak 1 at-61.6ppm, peak 2 at-64.4ppm, peak 3- 79.4ppm

Embodiment 8:Second eluting peak: 2-(5-(3-amino-5,6-two (trifluoromethyl) pyrazine-2-base)-1,3,4- Diazole-2-base) enantiomer 2 of-1,1,1-trifluoro propan-2-ol

1H NMR(400MHz,DMSO-d6):δ9.18-9.3(1H,br s),8.02-8.16(1H,br s),7.92(1H, s),1.88(3H,s).19F NMR (400MHz, DMSO-d6) peak 1 at-61.6ppm, peak 2 at-64.4ppm, peak 3- 79.4ppm。

Embodiment 9

2-(5-benzyl-1,3,4-diazole-2-base)-6-bromo-5-(trifluoromethyl) pyridine-3-amine

Step 1:The bromo-N'-of 3-amino-6-(2-phenyl acetyl)-5-(trifluoromethyl) pyridine-2-formylhydrazine

To stirred 3-amino-6-bromo-5-(trifluoromethyl) pyridine-2-formylhydrazine (intermediate H) (150mg, 0.502mmol) in the solution of NMP (4ml) add phenylacetic acid (57mg, 0.419mmol), be subsequently added DIPEA (366ul, 2.093mmol).This solution will be dividedly in some parts HATU (239mg, 0.628mmmol), and reactant mixture is stirred under RT 1h。

Reactant mixture is added in EtOAc (25ml), separate organic moiety, and with 0.1M HCl (2x15ml), water (25ml), the washing of 0.1M NaOH (2x15ml), water (25ml), saline (25ml), be dried (Mg SO4) and be also concentrated in vacuo.Will be thick Product is purified through silica gel chromatography 1:1EtOAc/ iso-hexane, obtains title compound, for yellow solid；1H NMR (400MHz,DMSO-d6)δ10.30(1H,s),10.20(1H,s),7.70(1H,s),7.38(4H,m),7.25(3H,m), 3.55(2H,s).LC-MS Rt=1.43min [M+H]+417.1 method 2minLC_v002.

Step 2:2-(5-benzyl-1,3,4-diazole-2-base)-6-bromo-5-(trifluoromethyl) pyridine-3-amine

To the stirred 3-bromo-N'-of amino-6-(2-phenyl acetyl)-5-(trifluoromethyl) pyridine-2-formylhydrazine (106mg, 0.254mmol) is at CHCl₃(2ml) suspension adds TEA (142ul, 1.016mmol), is subsequently added TsCl (145mg,0.762mmol).This mixture is heated to 65 DEG C and continues 3h.Solvent removed in vacuo, and by gained brown residue thing It is suspended in EtOAc (10ml).This mixture saline (10ml) is washed, is dried (MgSO4) and is concentrated in vacuo.By crude product Eluting with 85% isohexane/EtOAc through silica gel chromatography, obtain title compound, for yellow solid；

1H NMR(400MHz,DMSO-d6)δ7.85(1H,s),7.39(4H,m),7.30(3H,m),4.45(2H,s)。 LC-MS Rt=5.84min [M+H]+401.0 method 10minLC_v002.

According to method similar to Example 9 from 3-amino-6-bromo-5-(trifluoromethyl) pyridine-2-formylhydrazine (intermediate Or 3-amino-5-(trifluoromethyl) pyridine-2-formylhydrazine (intermediate compound I) and suitable processed with acid are for the embodiment (table in following table H) 2) compound.

Table 2

Embodiment 10

2-(5-(3-amino-6-cyclopropyl-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1, 1-trifluoro propan-2-ol

Step 1: the bromo-N'-of 3-amino-6-(3,3,3-tri-fluoro-2-hydroxy-2-methyl propiono)-5-(trifluoromethyl) pyrrole Pyridine-2-formylhydrazine

By cooling (0 DEG C) 3-amino-6-bromo-5-(trifluoromethyl) pyridine-2-formylhydrazine (intermediate H) (15g, 50.2mmol) the solution 3,3,3-tri-fluoro-2-hydroxy-2-methyl propanoic acid in anhydrous NMP (50ml) (8.72g, 55.2mmol), HATU (20.98g, 55.2mmol) process, go through subsequently 10 minutes be added dropwise over TEA (15.38ml, 110mmol).By this orange solution 0 DEG C of stirring, it is heated up to RT, and stirs 3 days.By reactant mixture at EtOAc (200mL) And distribute between 1M NaOH (200mL), shake and separate.Organic moiety saline (100mL) is washed, is dried (MgSO₄) and It is concentrated in vacuo, obtains orange.This grease is dissolved in methanol (50mL), and grinds with water (300mL), obtain title Compound, for yellow solid；

1H NMR(400MHz,DMSO-d6)δ10.3(1H,br s),10.08(1H,br s),7.72(1H,s),7.24 (2H,br s),7.16(1H,br s),1.54(3H,s).LC-MS Rt=1.04min [M+H]+439.0/441.1 method 2minLC_v003.19F NMR (400MHz, DMSO-d6) peak 1 is at-62.6ppm, and peak 2 is at-78ppm.

Step 2:The bromo-N'-of 3-amino-6-(3,3,3-tri-fluoro-2-methyl-2-(tri isopropyl silane base epoxide) propionyl Base)-5-(trifluoromethyl) pyridine-2-formylhydrazine

To the 3-bromo-N'-of amino-6-(3,3,3-tri-fluoro-2-hydroxy-2-methyl propiono)-5-(trifluoromethyl) pyridine-2- Formylhydrazine (step 1) (4g, 9.11mmol) adds triethylamine (1.270ml, 9.11mmol) in the suspension of DCM (60ml), Obtain yellow solution.This solution is cooled to 0 DEG C (ice bath), and with tri isopropyl silane base triflate (4.94ml, 18.22mmol) process.Reactant mixture is stirred 1 hour at 0 DEG C, is heated up to RT, and stirs 3 days.Reactant mixture is existed Distribute between DCM (100ml) and water (100ml), shake and separate each phase.Organic moiety saline (100ml) is washed, is dried And be concentrated in vacuo (MgSO4).By gained grease through silica gel chromatography 0-30%DCM eluant solution purification in isohexane, Obtain title compound；LC-MS Rt=1.58min [M+H]+595/597 method 2minLC_v003.

Step 3:The bromo-2-of 6-(5-(the fluoro-2-of 1,1,1-tri-(tri isopropyl silane base epoxide) acrylate-2-yl)-1,3,4-two Azoles-2-base)-5-(trifluoromethyl) pyridine-3-amine

(0 DEG C) bromo-N'-of 3-amino-6-(3,3,3-tri-fluoro-2-methyl-2-(tri isopropyl silane base epoxide) to cooling Propiono)-5-(trifluoromethyl) pyridine-2-formylhydrazine (5.42g, 9.10mmol) adds TEA in the solution of DCM (50ml) (3.81ml,27.3mmol).Gained solution is stirred 5 minutes at 0 DEG C, and with toluene sulfochloride (5.21g, 27.3mmol) place Reason.This mixture is heated up to room temperature, and stirs 48h.Reactant mixture is divided between DCM (100ml) and water (100ml) Join, shake and separate each phase.By organic moiety 1M NaOH (100ml), 1M HCl (100ml), water (100ml), saline (100ml) washing, is dried (MgSO4) (～10g) and is concentrated in vacuo, obtaining orange.By this grease through silica gel chromatography Purify with 0-15%EtOAc eluant solution in isohexane, obtain title compound；LC-MS Rt=7.41min [M+H]+ 577/579 method 10minLC_v003.

Step 4:6-cyclopropyl-2-(5-(the fluoro-2-of 1,1,1-tri-(tri isopropyl silane base epoxide) acrylate-2-yl)-1,3,4- Diazole-2-base)-5-(trifluoromethyl) pyridine-3-amine

At N₂In under RT to stirring in the bromo-2-of 6-(5-(the fluoro-2-of 1,1,1-tri-(tri isopropyl silane base epoxide) acrylate- 2-yl)-1,3,4-diazole-2-base)-5-(trifluoromethyl) pyridine-3-amine (step 3) (200mg, 0.346mmol) is anhydrous The solution of 1,4-dioxane (10ml) adds K₂CO₃(144mg, 1.039mmol), be subsequently added cyclopropylboronic acid (89mg, 1.039mmol) with palladium (IV) (40.0mg, 0.035mmol).By gained suspension stirring also heated at reflux (120 DEG C) overnight Continue 4 days.By this mixture warp(filter material) filters, and washs with water (30ml) and EtOAc (50ml).Separate filter Liquid, is dried (MgSO by organic moiety₄) and be concentrated in vacuo, obtaining title compound, it is without further purification for next step Suddenly；LC-MS Rt=7.75min [M+H]+539.6 method 10minLC_v003.

Step 5:2-(5-(3-amino-6-cyclopropyl-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2- Base)-1,1,1-trifluoro propan-2-ol

By 6-cyclopropyl-2-(5-(the fluoro-2-of 1,1,1-tri-(tri isopropyl silane base epoxide) acrylate-2-yl)-1,3,4-two Azoles-2-base)-5-(trifluoromethyl) pyridine-3-amine (step 4) (200mg, 0.371mmol) solution in anhydrous THF (10ml) The tetra-n-butyl ammonium fluoride (500mg, 0.750mmol) supported with silica gel processes.Gained redness suspension is stirred 5 points under RT Clock, and warp(filter material) filters.This mixture THF (10ml) is washed, and filter vacuum is concentrated, obtain orange Color grease.By this grease through silica gel chromatography 0-40%EtOAc eluant solution purification in isohexane, obtain titled Compound；

1H NMR(400MHz,DMSO-d6)δ7.74(1H,s),7.73(1H,s),6.92(2H,s),2.16(1H,m), 1.86 (3H, s), 1.01 (2H, m), 0.98 (2H, m) .19F NMR (400MHz, DMSO-d6) peak 1 is at-60.2ppm, peak 2- 79.6ppm.LC-MS Rt=1.23min [M+H]+383.2 method 2minLC_v003.

Embodiment 10.1

2-(5-(3-amino-6-methyl-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1- Trifluoro propan-2-ol

Title compound is prepared according to method similar to Example 10, with 2,4,6-trimethyl-1,3,5,2,4,6-tri- Oxa-three bora hexamethylene replaces cyclopropylboronic acid (step 4)；

1H NMR (400MHz, the DMSO-d6) δ 7.79 of thick raceme material (1H, s), 7.75 (1H, s), 7.0 (2H, s),2.53(3H,s),1.89(3H,s).LC-MS Rt=3.75min [M+H]+357.2 method 10minLC_v003.Outside Cu 19F NMR (400MHz, the DMSO-d6) peak 1 of racemic material is at-61.9ppm, and peak 2 is at-79.6ppm.

Embodiment 10.2

2-(5-(3-amino-6-bromo-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-three Fluorine propan-2-ol

Title compound is prepared, with the bromo-2-of 6-(5-(1,1,1-tri-fluoro-2-(three according to method similar to Example 10 Isopropyl silylation epoxide) acrylate-2-yl)-1,3,4-diazole-2-base)-5-(trifluoromethyl) pyridine-3-amine (step 3) replacement 6-cyclopropyl-2-(5-(the fluoro-2-of 1,1,1-tri-(tri isopropyl silane base epoxide) acrylate-2-yl)-1,3,4-diazole-2-base)- 5-(trifluoromethyl) pyridine-3-amine (step 5)；1H NMR(400MHz,DMSO-d6)δ7.90(1H,s),7.84(1H,s), 7.34(2H,s),1.86(3H,s).LC-MS Rt=4.42min [M+H]+423 method 10minLC_v003.

Embodiment 10.3

2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoropropyl- 2-alcohol

Step 1:2-(5-(the fluoro-2-of 1,1,1-tri-(tri isopropyl silane base epoxide) acrylate-2-yl)-1,3,4-diazole-2- Base)-5-(trifluoromethyl) pyridine-3-amine

According to 2-bromo-with 6-(5-(the fluoro-2-of 1,1,1-tri-(tri isopropyl silane base epoxide) acrylate-2-yl)-1,3,4-two Azoles-2-base) the similar method of-5-(trifluoromethyl) pyridine-3-amine (embodiment 10, step 3) prepares title compound, uses 3-ammonia Base-5-(trifluoromethyl) pyridine-2-formylhydrazine (intermediate compound I) replaces 3-amino-6-bromo-5-(trifluoromethyl) pyridine-2-formyl Hydrazine (intermediate H) (step 1)；

LCMS:Rt=1.66min；[M+H]+499.3 method 2minLC_v003.

Step 2:2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-three Fluorine propan-2-ol

According to 2-(5-(3-amino-6-cyclopropyl-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2- Base) the similar method of-1,1,1-trifluoro propan-2-ol (embodiment 10 step 5), from 2-(5-(1,1,1-tri-fluoro-2-(triisopropyl Silylation epoxide) acrylate-2-yl)-1,3,4-diazole-2-base)-5-(trifluoromethyl) pyridine-3-amine (step 1) prepares titled Compound；1H NMR(400MHz,DMSO-d6)δ8.31(1H,s),7.78(1H,s),7.73(1H,s),7.21(2H,s),1.86 (3H,s).LCMS:Rt=1.07min；[M+H]+343.1 method 2minLC_v003.

Embodiment 10.4 and 10.5

By 2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-under the conditions of using supercritical fluid chromatography below Base)-1,3,4-diazole-2-base)-1,1,1-trifluoro propan-2-ol (embodiment 10.3) chiral separation prepares following compound:

Flowing phase: 10% isopropanol+0.1%DEA/90%CO₂

Post: Chiralpak AS-H, 250x10mm, 5 μm

Detection: UV@220nm

Flow velocity: 10ml/min

Sample concentration: 118mg is in 1.25ml EtOH.

Embodiment 10.4:First eluting peak.2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4- Diazole-2-base) enantiomer 1 of-1,1,1-trifluoro propan-2-ol

1H NMR(400MHz,DMSO-d6)δ8.31(1H,s),7.78(1H,s),7.73(1H,s),7.21(2H,s), 1.86(3H,s).LC-MS Rt=3.36min [M+H]+343.4 method 10minLC_v003.

Embodiment 10.5:Second eluting peak.2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4- Diazole-2-base) enantiomer 2 of-1,1,1-trifluoro propan-2-ol

1H NMR(400MHz,DMSO-d6)δ8.31(1H,s),7.78(1H,s),7.73(1H,s),7.21(2H,s), 1.86(3H,s).LC-MS Rt=3.35min [M+H]+343.4 method 10minLC_v003.

Embodiment 10.6

2-(5-(3-amino-6-bromo-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base) propan-2-ol

According to the method similar with embodiment 10 (leaving out step 4), from 3-amino-6-bromo-5-(trifluoromethyl) pyridine-2- Title compound prepared by formylhydrazine (intermediate H) and 2-hydroxy-2-methyl propanoic acid；

1H NMR(400MHz,DMSO-d6)δ7.9(1H,s),7.3(2H,s),6.0(1H,s),1.6(6H,s)。

LC-MS Rt=1.07min [M+H]+367.1 method 2minLC_v003.

Embodiment 11

(R)-2-(5-(amino (phenyl) methyl)-1,3,4-diazole-2-base)-6-bromo-5-(trifluoromethyl) pyridine-3- Amine

Step 1:(R)-2-(2-(3-amino-6-bromo-5-(trifluoromethyl) pyridine-2-formoxyl) diazanyl)-2-oxo-1- Phenylethylcarbamate benzyl chloroformate

By 3-amino-6-bromo-5-trifluoromethylpyridin-2-formic acid (intermediate A) (200mg, 0.702mmol) at NMP Suspension in (6ml) (R)-2-diazanyl-2-oxo-1-phenylethylcarbamate benzyl chloroformate (intermediate JR) (231mg, 0.772mg) process, be dividedly in some parts HATU (293mg, 0.772mmol) subsequently.After stirring 45 minutes under RT, reaction is mixed Thing adds in EtOAc (50ml), and washs with 0.1M NaOH (50ml).Water layer is extracted with EtOAc (25ml) again.To merge Organic moiety use water (75ml), saline (50ml) washing, be dried (MgSO₄) and be concentrated in vacuo.Crude product is dissolved in MeOH, on Sample is to pre-wetted (MeOH)PE-AX (anion exchange) post.By this post MeOH (70ml) eluting, and will filter Liquid is concentrated in vacuo, and obtains yellow solid, is placed under fine vacuum, obtains title compound, for yellow solid；

1H NMR (400MHz, DMSO-d6) δ 10.50 (2H, wide), 8.04 (1H, d), 7.70 (1H, s), 7.52 (2H, d), 7.35(8H),7.21(2H,s),5.45(1H,d),5.06(2H).There is some NMP and acetic acid.LC-MS Rt=1.55min [M + H]+568.1 methods 2minLC_v002.

Step 2:(R)-(5-(3-amino-6-bromo-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base) (phenyl) methyl carbamic acid benzyl ester

To (R)-2-(2-(3-amino-6-bromo-5-(trifluoromethyl) pyridine-2-formoxyl) diazanyl)-2-oxo-1-phenyl Ethyl carbamic acid benzyl ester (step 1) (360mg, 0.636mmol) is at CHCl₃(6ml) solution adds triethylamine (354ul, 2.54mmol), is subsequently added toluene sulfochloride (242mg, 1.271mmol).By gained mixture 65 DEG C of stirrings 2h, and be concentrated in vacuo subsequently.This residue is dissolved in EtOAc (20ml), and washs with saline (20ml), be dried (MgSO₄), And be evaporated under reduced pressure.By eluting for crude on silica gel chromatographic grade 75% isohexane/EtOAc, obtain title compound；1H NMR (400MHz,DMSO-d6)δ8.85(1H,d),7.85(1H,s),7.50(2H,d),7.45-7.30(10H),6.32(1H,d), 5.10(2H,s).LC-MS Rt=1.64min [M+H]+550 method 2minLC_v002.

Step 3:(R)-2-(5-(amino (phenyl) methyl)-1,3,4-diazole-2-base) the bromo-5-of-6-(trifluoromethyl) Pyridine-3-amine

To stirred (R)-(5-(3-amino-6-bromo-5-(trifluoromethyl) pyridine-2-base)-1,3,4-in nitrogen Diazole-2-base) (phenyl) methyl carbamic acid benzyl ester (step 2) (55mg, 0.100mmol) is at the solution of anhydrous DCM (6ml) Middle addition Iodotrimethylsilane (55ul, 0.401mmol).Reactant mixture is stirred overnight under RT.Use SPE post (SCX-2) eluting with the MeOH solution (5ml) of 3.5M ammonia subsequently with MeOH, obtain title compound；1H NMR(400MHz,DMSO-d6)δ7.85(1H,s),7.50(2H,d),7.49(2H,t),7.31(5H),5.49(1H,s)。LC- MS Rt=1.39min [M+H]+416.1 method 2minLC_v002.

Embodiment 12.0

2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4-thiadiazoles-2-base)-1,1,1-trifluoropropyl- 2-alcohol

Step 1:3-amino-N'-(3,3,3-tri-fluoro-2-hydroxy-2-methyl propiono)-5-(trifluoromethyl) pyridine-2- Formylhydrazine

Add in the solution of NMP (5ml) to 3,3,3-tri-fluoro-2-hydroxy-2-methyl propanoic acid (324mg, 2.049mmol) HATU (935mg, 2.458mmol) and DIPEA (1.073ml, 6.15mmol).After stirring 5 minutes, add 3-amino-5-(trifluoro Methyl) pyridine-2-formylhydrazine (intermediate compound I) (451mg, 2.049mmol), and under RT, continue stirring 1h.By reactant mixture Dilute with water, and extract with EtOAc.By the organic extract 1M HCl (100ml) merged, water (2x100ml), salt washing Wash, be dried (phase separator) and be concentrated in vacuo.Through silica gel chromatography 0-30%EtOAc eluant solution purification in isohexane, Obtain title compound；1H NMR400MHz,DMSO-d6:δ10.3(1H,s),10.0(1H,s),8.0(1H,s),7.5(1H, S), 7.1 (3H, s, overlapping), 1.5 (3H, s).LC-MS Rt=0.98min [M+H]+361.2 method 2minLC_v003.

Step 2:2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4-thiadiazoles-2-base)-1,1,1-three Fluorine propan-2-ol

3-amino-N'-(3,3,3-tri-fluoro-2-hydroxy-2-methyl propiono)-5-(trifluoromethyl) pyridine-2-will be comprised The formylhydrazine (184mg, 0.511mmol) mixture in toluene (5.108ml) lawesson reagent (310mg, 0.766mmol) place Reason, and it is heated at reflux 1h.By reactant mixture dilute with water, and extract with EtOAc.By the organic extract saturated carbon of merging Acid hydrogen sodium solution, saline washing, be dried (phase separator) and be concentrated in vacuo.Through silica gel chromatography 0-30%EtOAc at isohexane In eluant solution purification, obtain orange jelly.30-70%MeCN aqueous solution is used further through preparative reversed-phase HPLC (0.1%TFA) eluting, obtain title compound；1H NMR(400MHz,DMSO-d6)δ8.2(1H,s),8.0(1H,s), 7.6(1H,s),7.4(2H,s),1.9(3H,s).LC-MS Rt=4.03min [M+H]+359.4 method 10minLC_v003.

Embodiment 13.0

(R)-2-[5-(3-amino-4-chloro-6-methoxyl group-5-trifluoromethylpyridin-2-base)-[1,3,4] diazole-2- Base]-1,1,1-trifluoro-propyl-2-alcohol

To (R)-2-(5-(3-amino-6-methoxyl group-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2- Base) to add trichlorine in the solution of acetonitrile (1ml) different for-1,1,1-trifluoro propan-2-ol (embodiment 2) (150mg, 0.403mmol) Cyanuric acid (37.5mg, 0.161mmol), and gained mixture microwave radiation is heated 30 minutes at 100 DEG C, exist subsequently 125 DEG C are heated 10 minutes.This mixture is distributed between EtOAc and water.Separate organic moiety, and use saturated NaHCO₃Molten Liquid, saline wash, and are dried (MgSO₄) and be concentrated in vacuo.By crude on silica gel chromatographic grade 0-30%EtOAc in isohexane Eluant solution purification, obtains title compound；

LC-MS Rt=4.40min [M+H]+407.1 method 10minLC_v003.

1H NMR(400MHz,DMSO-d6)δ7.80(1H,s),6.90(2H,s),3.95(3H,s),1.87(3H,s)

Embodiment 13.1

(S)-2-(5-(3-amino-4-chloro-6-methoxyl group-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2- Base)-1,1,1-trifluoro propan-2-ol

Prepare title compound according to the method similar with embodiment 13.0, with (S)-2-(5-(3-amino-6-methoxyl group- 5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoro propan-2-ol (embodiment 3) replacement (R)- 2-(5-(3-amino-6-methoxyl group-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoropropyl- 2-alcohol (embodiment 2)；

LC-MS Rt=1.25min [M+H]+407.1 method 2minLC_v003.

1H NMR(400MHz,DMSO-d6)δ7.81(1H,s),6.90(2H,s),3.96(3H,s),1.88,(3H,s)。

Embodiment 14.0

(S)-2-[5-(3-amino-4-ethyl-6-methoxyl group-5-trifluoromethylpyridin-2-base)-[1,3,4] diazole- 2-yl]-1,1,1-trifluoro-propyl-2-alcohol

Step 1: (S)-2-(5-(3-amino-6-methoxyl group-5-(trifluoromethyl)-4-vinylpridine-2-base)-1,3, 4-diazole-2-base)-1,1,1-trifluoro propan-2-ol

To (S)-2-(5-(3-amino-4-chloro-6-methoxyl group-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole- 2-yl)-1,1,1-trifluoro propan-2-ol (embodiment 13.1) (160mg, 0.393mmol) adds two in the solution of MeCN (1ml) (triphenylphosphine) Palladous chloride. (II) (Aldrich) (83mg, 0.118mmol), is subsequently added 4,4,5,5-tetramethyl-2-ethylene Base-1,3,2-dioxaborolan alkane (Sigma Aldrich) (0.087ml, 0.511mmol).Add 2M sodium carbonate (0.885ml, 1.770mmol), and gained mixture microwave radiation is heated 30 minutes at 130 DEG C.By this mixture warpFilter, and wash with EtOAc.Filtrate is diluted with EtOAc (50ml) further, and uses saturated NaHCO₃Solution, Water, saline wash, and use MgSO₄It is dried.Add Isolute Si-TMT (2,4,6-tri-thiol triazine silica gel, palladium scavenger), This mixture is stirred 30 minutes and filters.Solvent removed in vacuo, obtains title compound, and it the most directly makes With；

LC-MS Rt=1.25min [M+H]+399.3 method 2minLC_v003.

Step 2:(S)-2-[5-(3-amino-4-ethyl-6-methoxyl group-5-trifluoromethylpyridin-2-base)-[1,3,4] Diazole-2-base]-1,1,1-trifluoro-propyl-2-alcohol

To stirring in (S)-2-(5-(3-amino-6-methoxyl group-5-(trifluoromethyl)-4-vinylpridine-2-base)- 1,3,4-diazole-2-base)-1,1,1-trifluoro propan-2-ol (step 1) (153mg, 0.384mmol) is molten EtOH's (10ml) Liquid adds palladium dydroxide/carbon (43.2mg, 0.307mmol), is subsequently added ammonium formate (969mg, 15.37mmol).Will reaction Mixture is heated at reflux 30 minutes.By this mixture warpFilter, and dilute with EtOAc.By filtrate with saturated NaHCO₃Solution, water, saline wash, and use MgSO₄It is dried.Addition Isolute Si-TMT (2,4,6-tri-thiol triazine silica gel, Palladium scavenger), and this mixture is stirred 30 minutes and filters.Solvent removed in vacuo, and by crude on silica gel chromatographic grade 0- 30%EtOAc eluant solution purification in isohexane, obtains title compound；

LC-MS Rt=4.49min [M+H]+401.2 method 10minLC_v003.

1H NMR(400MHz,DMSO-d6)δ7.76(1H,s),6.72(2H,s),3.91(3H,s),2.85(2H,m), 1.87(3H,s),1.17(3H,t)。

Embodiment 14.1

(S)-2-[5-(3-amino-4-isopropenyl-6-methoxyl group-5-trifluoromethylpyridin-2-base)-[1,3,4] Diazole-2-base]-1,1,1-trifluoro-propyl-2-alcohol

According to (S)-2-(5-(3-amino-6-methoxyl group-5-(trifluoromethyl)-4-vinylpridine-2-base)-1,3, 4-diazole-2-base) the similar method of-1,1,1-trifluoro propan-2-ol (embodiment 14.0 step 1) prepares title compound, uses 4,4,5,5-tetramethyl-2-(acrylate-1-alkene-2-base)-1,3,2-dioxaborolan alkane replaces 4,4,5,5-tetramethyl-2-second Thiazolinyl-1,3,2-dioxaborolan alkane；

LC-MS Rt=4.80min [M+H]+413.2 method 10minLC_v003.

1H NMR(400MHz,DMSO-d6)δ7.77(1H,s),6.33(2H,s),5.48(1H,s),5.00(1H,s), 3.94(3H,s),2.01(3H,s),1.87(3H,s)。

Embodiment 14.2

(S)-2-[5-(3-amino-4-isopropyl-6-methoxyl group-5-trifluoromethylpyridin-2-base)-[1,3,4] two Azoles-2-base]-1,1,1-trifluoro-propyl-2-alcohol

According to (S)-2-[5-(3-amino-4-ethyl-6-methoxyl group-5-trifluoromethylpyridin-2-base)-[1,3,4] Diazole-2-base]-1, the method that 1,1-trifluoro-propyl-2-alcohol (embodiment 14.0 step 2) is similar, from (S)-2-[5-(3-ammonia Base-4-isopropenyl-6-methoxyl group-5-trifluoromethylpyridin-2-base)-[1,3,4] diazole-2-base]-1,1,1-three is fluoro- Propan-2-ol (embodiment 14.1) prepares title compound；

LC-MS Rt=4.95min [M+H]+415.2 method 10minLC_v003.

1H NMR(400MHz,DMSO-d6)δ7.76(1H,s),6.51(2H,s),3.90(3H,s),3.60(1H,m), 1.87(3H,s),1.41(6H,d)。

Embodiment 14.3

(S)-2-[5-(3-amino-6-methoxyl group-4-phenyl-5-trifluoromethylpyridin-2-base)-[1,3,4] diazole- 2-yl]-1,1,1-trifluoro-propyl-2-alcohol

According to (S)-2-(5-(3-amino-6-methoxyl group-5-(trifluoromethyl)-4-vinylpridine-2-base)-1,3, 4-diazole-2-base) the similar method of-1,1,1-trifluoro propan-2-ol (embodiment 14.0 step 1) prepares title compound, uses 4,4,5,5-tetramethyl-2-phenyl-1,3,2-dioxaborolan alkane replaces 4,4,5,5-tetramethyl-2-vinyl-1,3, 2-dioxaborolan alkane；

LC-MS Rt=4.98min [M+H]+449.3 method 10minLC_v003.

1H NMR(400MHz,DMSO-d6)δ7.79(1H,s),7.56(3H,m),7.30(2H,m),5.80(2H,s), 3.99(3H,s),1.88(3H,s)

Prepared by intermediate

Intermediate A

3-amino-6-bromo-5-trifluoromethylpyridin-2-formic acid

Intermediate A 1:2-bromo-3-nitro-5-trifluoromethylpyridin

3-nitro-5-(trifluoromethyl) pyridine-2-alcohol (31.00g, 149mmol) is dissolved in acetonitrile (250ml), obtains Deep brown solution.Add phosphorus oxybromide (V) (85g, 298mmol), and this mixture is heated at reflux 4 hours, and subsequently at RT Under be stirred overnight.Reactant mixture is poured into quencher in the water (600ml) comprising sodium bicarbonate (110g) being stirred vigorously.Should Dark brown mixture DCM extracts (3x200ml), and organic phase washed with water (200ml) and saline (100ml) is washed, and is dried (MgSO₄) and be concentrated in vacuo, obtain title product, for brown oil.¹H-NMR:[400MHz,CDCl₃,δ8.87(1H,d,J =1.4Hz, ArH), 8.39 (1H, d, J=1.9Hz, ArH).

Intermediate A 2:3-nitro-5-trifluoromethylpyridin-2-formonitrile HCN

Bromo-for 2-3-nitro-5-trifluoromethylpyridin (10.00g, 36.87mmol) is under agitation dissolved in toluene (250ml), in, pale yellow solution is obtained.Add tetrabutyl ammonium bromide (11.90g, 36.9mmol), be subsequently added Cupricin. (I) (9.92g, 111mmol), and this mixture is heated at reflux 9 hours.After being cooled to RT, by reactant mixture at water (750ml) and distribute between EtOAc (750ml).Organic moiety is merged, washs with water (2x250ml) and saline (100ml), It is dried (MgSO₄) and be concentrated in vacuo, obtain title product.¹H-NMR:[400MHz,DMSO-d₆]δ9.55(1H,m,ArH),9.24 (1H,m,ArH)

Intermediate A 3:3-amino-5-trifluoromethylpyridin-2-methyl formate

3-nitro-5-trifluoromethylpyridin-2-formonitrile HCN (6.5g, 29.9mmol) is dissolved in EtOAc (150ml), obtains Pale yellow solution.Add the palladium (3.19g, 2.99mmol) on 10% activated carbon, and reactant mixture is stirred in hydrogen 18 Hour.Reactant mixture is filtered, and is concentrated in vacuo.Thick residue is dissolved in dense HCl (45ml), and it is little to be heated at reflux 24 Time.Reactant mixture is cooled to RT, and is concentrated in vacuo.This solid is dissolved in MeOH (200ml), and adds sulphuric acid (8ml). Gained solution is heated at reflux 84 hours.This reactant is cooled to RT, subsequently by adding 10%NaHCO_3(aq)(600ml) in With.Product is extracted in DCM (3x200mL), and by the organic phase washed with water (200ml) merged, saline (50mL) washing, (MgSO₄) and be concentrated in vacuo.By gained solid through silica gel chromatography: gradient elution: isohexane (500ml), 10%EtOAc exist Solution (1000mL) in isohexane, 20%EtOAc solution (1500mL) in isohexane, obtain title compound, for shallow Yellow solid.¹H-NMR:[400MHz,DMSO-d₆] δ 8.13 (1H, d, J=1.7Hz, ArH), 7.60 (1H, d, J=1.3Hz, ArH),7.01(2H,br,NH₂),3.85(3H,s,ArOCH₃),m/z221.1[M+H]⁺

Intermediate A 4:3-amino-6-bromo-5-trifluoromethylpyridin-2-methyl formate

3-amino-5-trifluoromethylpyridin-2-methyl formate (9.49g, 43.16mmol) is dissolved in water (300mL). Add sulphuric acid (4.60mL, 86mmol), go through subsequently 30 minutes and be added dropwise over bromine (2.222mL, 43.1mmol) at acetic acid Solution in (29.6mL, 517mmol).Reactant mixture is stirred 18 hours under RT.Add 100ml water, add the most again Enter the bromine/AcOH mixture (550 μ L bromines are in 7.4mL AcOH) of 0.25 equivalent, and reactant mixture is stirred for 90 under RT Minute.Reactant mixture 500mL water is diluted, and by adding solid NaHCO₃(～85g) neutralizes.This suspension is used DCM (3x300mL) extracts, and the saturated NaHCO of organic facies that will merge_3(aq)(250mL), water (250mL) and saline (100mL) Washing, is dried (MgSO₄) and be concentrated in vacuo.By this thick material recrystallization from the MeOH (～300mL) of boiling, obtain title and produce Thing, for orange solid m/z301.0 [M+H]⁺¹H-NMR:[400MHz,DMSO-d₆]δ7.77(1H,s,ArH),7.17(2H,s, NH₂),3.86(3H,s,ArCO₂CH₃)。

Intermediate A:3-amino-6-bromo-5-trifluoromethylpyridin-2-formic acid

3-amino-6-bromo-5-trifluoromethylpyridin-2-methyl formate (1.40g, 4.68mmol) is suspended in MeOH (15mL) in；Add sodium hydroxide (2.0M aqueous solution) (14.04mL, 28.1mmol), and by this suspension under RT stirred Night.Reactant mixture is concentrated in vacuo, and gained residue is dissolved in water (100mL), subsequently by adding 5.0M HCl (aq) acidifying.This product is extracted in ethyl acetate (2x75mL), and by extract use water (50mL) merged, saline (25mL) washing, is dried (MgSO₄) and be concentrated in vacuo, obtain title product, for yellow solid.¹H-NMR:[400MHz,DMSO- d₆]δ13.24(1H,br s,CO₂H),7.74(1H,s,ArH),7.1792H,br s ArNH₂)。m/z285.1,287.1[M+H ]⁺

Intermediate B

3-amino-6-methoxyl group-5-trifluoromethylpyridin-2-formic acid

Intermediate B 1:The bromo-3-of 6-(2,5-Dimethyl-pyrrol-1-base)-5-trifluoromethylpyridin-2-methyl formate

3-amino-6-bromo-5-trifluoromethylpyridin-2-methyl formate (intermediate A 4) (2g, 6.69mmol) is suspended in In toluene (8ml), be subsequently added p-methyl benzenesulfonic acid (TsOH) (0.115g, 0.669mmol) and acetonyl acetone (0.941ml, 8.03mmol).Reactant mixture is heated at reflux 2 hours, and is cooled to RT overnight.By dense for dark red for gained/dark solution vacuum Contracting is to remove toluene, and by this thick residue 200ml EtOAc dilution, uses NaHCO₃(50ml) washing, is dried (MgSO₄) And be concentrated in vacuo, obtain brown solid；LC-MS Rt=5.58min [M+H]+377/379 (method 10minLC_v002).

1H NMR(400MHz,DMSO-d6)δ8.50(1H,s),7.77(2H,s),5.83(3H,s),1.90(6H,s)； 19F NMR(400MHz,DMSO-d6)δ-62.26(CF3,s)

Intermediate B 2:3-(2,5-Dimethyl-pyrrol-1-base)-6-methoxyl group-5-trifluoromethylpyridin-2-formic acid

By bromo-for 6-3-(2,5-Dimethyl-pyrrol-1-base)-5-trifluoromethylpyridin-2-methyl formate (2g, 5.30mmol) it is dissolved in MeOH (40ml), and processes with 2M NaOH (20ml), obtain suspension, it is stirred 1 hour under RT, Obtain settled solution.Solvent removed in vacuo, and residue 5M HCl is acidified to pH1.By this mixture EtOAc (200ml) extraction, and organic extract is dried (MgSO₄) and be concentrated in vacuo, obtain title compound, for dark brown solid, It without further purification and is directly used in next step；LC_MS Rt=1.50min [M+H]+315.2.1/316.2 (method 2minLC_v002)；1H NMR(400MHz,DMSO-d6)δ14.42-12.61(COOH,b hump),8.25(1H,s),5.84 (2H,s),4.13(3H,s),1.97(6H,s)；19FNMR(400MHz,DMSO-d6)δ-62.43(CF3,s).

Intermediate B:3-amino-6-methoxyl group-5-trifluoromethylpyridin-2-formic acid

By 3-(2,5-Dimethyl-pyrrol-1-base)-6-methoxyl group-5-trifluoromethylpyridin-2-formic acid (833mg, 2.65mmol) it is dissolved in EtOH (45ml) and water (23ml).TEA (1.102ml, 7.95mmol) is added in this mixture, with Rear addition oxammonium hydrochloride. (1842mg, 26.5mmol).By gained mixture heated at reflux overnight.After being cooled to RT, by this mixing Thing and 20gPE-AX (for separate acid compound the quaternary ammonium functional group with chemical bonding based on Silicon stone Adsorbent) stir 30 minutes, with MeOH (100ml), 1M HCl:MeCN2:8 (200ml) wash.Organic moiety is removed, and This mixture is filtered.Filtrate is acidified with 2M HCl (50ml), and vacuum removes EtOH.By aqueous fractions with DCM (200ml) Extraction, and organic extract is dried (MgSO4) and is concentrated in vacuo, obtain brown oil.Through silica gel chromatography DCM:MeOH Eluting, obtain title compound, for yellow solid: LC-MS Rt=2.90min [M+H]+237 (method 10minLC_ v002)

1H NMR (400MHz, DMSO-d6) δ 9.62-7.79 (NH2, wide hump (b hump)), 7.70 (1H, s), 3.89 (3H,s)；19F NMR(400MHz,DMSO-d6)δ-62.92(CF3,s).

Intermediate C

2-(hydrazine carbonyl)-6-methoxyl group-5-(trifluoromethyl) pyridin-3-yl carbamate

Intermediate C1:3-amino-6-methoxyl group-5-(trifluoromethyl) pyridine-2-methyl formate

By 3-amino-6-methoxyl group-5-trifluoromethylpyridin-2-formic acid (intermediate B) 5.5g, 23.29mmol) it is dissolved in In MeOH (90ml).It is added dropwise over H₂SO₄(6.21ml, 116mmol), and this solution is heated to backflow lasting 4 hours.Will be anti- Answer mixture to be concentrated in vacuo to about 15ml, and add water (15ml).By being carefully added into solid NaHCO₃PH is adjusted to pH9. Add water (100ml), and this mixture DCM is extracted.The organic extract of merging is concentrated in vacuo, obtains title compound Thing.

Intermediate C2:3-(tertbutyloxycarbonylamino)-6-methoxyl group-5-(trifluoromethyl) pyridine-2-methyl formate

To stirring in 3-amino-6-methoxyl group-5-(trifluoromethyl) pyridine-2-methyl formate (step 1) (4.989g, 19.94mmol) in the solution of DCM (100ml) add DIPEA (2.84g, 21.94mmol), Boc acid anhydride (4.79g, 21.94mmol), DMAP (2.436g, 19.94mmol) it is subsequently added.After stirring 3 hours, wash reactant mixture with water (3 Secondary), it is dried (MgSO₄) and be concentrated in vacuo.Crude on silica gel chromatographic grade 0%-10% isohexane: EtOAc is eluting, obtains Title compound.

Intermediate C:2-(hydrazine carbonyl)-6-methoxyl group-5-(trifluoromethyl) pyridin-3-yl carbamate

By 3-(tertbutyloxycarbonylamino)-6-methoxyl group-5-(trifluoromethyl) pyridine-2-methyl formate (step 2) (3.1g, 8.85mmol) suspension monohydrate hydrazine (1.108g, 22.12mmol) in anhydrous MeOH (20ml) processes, and By this suspension heated overnight under reflux.By this mixture dilute with water, and gained precipitation is collected by filtration, and in vacuum Stove is dried, obtains title compound, for brown solid (3.01g).LC-MS Rt=1.27min [M+H]+251 [-cut Boc group] (method HighpH_v003.).

Intermediate D

The fluoro-2 Methylpropionic acid of 2-(benzyloxy)-3,3,3-three

Intermediate D1:3,3,3-tri-fluoro-2-hydroxy-2-methyl propanoic acid benzyl ester

Fluoro-for 3,3,3-tri-2-hydroxy-2-methyl propanoic acid (0.6g, 3.80mmol) is dissolved in MeCN (5ml).Add DIPEA (0.663ml, 3.80mmol), and stir 5 minutes.Add benzyl bromide a-bromotoluene (541mg, 3.16mmol), and by reactant mixture Stir under RT 16 hours, stir 16 hours at 70 DEG C subsequently.After being cooled to RT, solvent removed in vacuo, and by gained residue It is dissolved in DCM.This mixture is washed with water, and by phase separator separation organic moiety.Solvent removed in vacuo, obtains title Compound.

Intermediate D2:The fluoro-2 Methylpropionic acid benzyl ester of 2-(benzyloxy)-3,3,3-three

By cooling (0 DEG C) 3,3,3-tri-fluoro-2-hydroxy-2-methyl propanoic acid benzyl ester (intermediate D1) (100mg, 0.403mmol) treated with sodium hydride (16.11mg, 0.403mmol) in DMF (4ml) processes, and uses benzyl bromide a-bromotoluene subsequently (0.048ml, 0.403mmol) processes, and stirs 2 hours at 0 DEG C.Reactant mixture is heated up to room temperature, and is further continued for stirring Mix 3 hours.Reactant mixture is distributed between EtOAc and 0.1M HCl solution.Organic facies saturated brine is washed, uses sulfur Acid magnesium is dried, and is concentrated in vacuo, and obtains title compound, for grease.

Intermediate D3:The fluoro-2 Methylpropionic acid of 2-(benzyloxy)-3,3,3-three

Fluoro-for 2-(benzyloxy)-3,3,3-three 2 Methylpropionic acid benzyl ester (intermediate D3) (170mg, 0.502mmol) is existed 2M sodium hydride (0.502ml, 1.005mmol) the 2M process of solution in MeOH (5ml), and stir 2 hours under RT.Vacuum is removed Remove methanol, and residue is soluble in water, and wash with EtOAc.Aqueous phase 5M HCl is acidified, and extracts with EtOAc.To close And organic extract with saturated brine wash, be dried with magnesium sulfate, and be concentrated in vacuo, obtain title product, for defecation；¹H NMR(400MHz,DMSO–d6)δ14.08(1H,br s),7.35(5H,m),4.62(2H,dd),1.64(3H,s)。

Intermediate E

3-amino-5-trifluoromethyl-pyrazine-2-Ethyl formate

Intermediate E 1:Amidino-nitroso-group-ethyl acetate

Go through in the ammonia of 2M solution in EtOH (152ml, 0.304mmol) 30 minutes at 0 DEG C to 5 DEG C and add second Epoxide carbonyl iminoester hydrochloride (25g, 0.127mmol).Reactant is stirred vigorously at such a temperature 3 hours, so After once add Cyanogran. solution (9.63g, 0.139mmol) in water.By adding 5N HCl, the pH of this mixture is adjusted To pH6.Reactant mixture is stirred overnight under RT.The yellow mercury oxide that filtered under vacuum is formed, washes with water and is dried, obtain Title compound；

¹H NMR(400MHz,DMSO–d6)δ10.1(2H,br s),7.6(2H,br s),4.3(2H,q),1.3(3H, t)。

Intermediate E 2:Amino-amidino-ethyl acetate

To amidino-nitroso-group-ethyl acetate (5.5g, 31.4mmol) ethanol/5M HCl's (1:1 ratio, 250ml) Solution adds 10%Pd/C (1.3g).Reactant mixture is under low pressure hydrogenated (H_2(g)) 2 nights.Warp(filter material) is filtered Except Pd/C, and filter vacuum is concentrated, obtain title compound, for white solid.It is used for next step with thick material.

Intermediate E:3-amino-5-trifluoromethyl-pyrazine-2-Ethyl formate

20% trifluoropropyl is added in the mixture of amino-amidino-ethyl acetate (2g, 9.22mmol) and water (50ml) Keto-aldehyde (2.32g, 18.43mmol) aqueous solution.Sodium acetate (5.29g, 64.52mmol) (reaction mixing is added in this mixture The pH of thing is pH5).This reactant mixture is stirred overnight under RT.Gained precipitates vacuum filter, use dissident through silica gel chromatography Alkane: EtOAc (0-10%EtOAc gradient) is eluting, obtains title compound.

¹H NMR(400MHz,DMSO–d6)δ8.4(1H,s),7.8(2H,br s),4.4(2H,q),1.4(3H,t)。

Intermediate F

3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid

Intermediate F1:3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-Ethyl formate

To 3-amino-5-trifluoromethyl-pyrazine-2-Ethyl formate (intermediate E) (30mg, 0.13mmol) at acetic acid (5ml) solution adds sodium carbonate (15mg, 0.14mmol).In this mixture, add half content bromine (7 μ L, 0.13mmol) the solution in acetic acid (5ml), is subsequently added sodium carbonate ((15mg, 0.14mmol).Add residue bromine at acetic acid In solution, and reactant mixture is stirred 2 hours under RT.By this mixture dilute with water, and by true for gained yellow mercury oxide Empty filtration, obtains title compound.

Intermediate F:3-amino-6-bromo-5-trifluoromethyl-pyrazine-2-formic acid

3-amino-5-trifluoromethyl-pyrazine-2-Ethyl formate (10g, 31.8mmol) in stirring is at ethanol (20ml) Solution in add 2M NaOH (20ml, 31.8mmol).

Gained solution is stirred 5 minutes under RT, and pours in water (50ml).By adding 1M HCl, pH is adjusted to pH6. Gained suspension vacuum is filtered, washs with water (20ml) and be dried, obtain title compound；

¹H NMR(400MHz,DMSO–d6)δ7.98(2H,s)。

Intermediate G

3-amino-5,6-two (trifluoromethyl) pyrazine-2-formylhydrazine

Intermediate G1:3-amino-5,6-two (trifluoromethyl) pyrazine-2-urethanes

Add in the solution of DMF (5ml) to amino-amidino-ethyl acetate (intermediate E 2) (1.25g, 8.61mmol) Enter 1,1, Isosorbide-5-Nitrae, 4,4-hexafluoro butane-2,3-diketone (5g, 25.8mmol), and this mixture is stirred 25 days at RT.By this Huang Color contamination suspension distributes between EtOAc (50ml) and water (50ml), and is washed by this organic moiety saline (30ml), is dried (MgSO₄) and be concentrated in vacuo.Crude product is eluting through the LC-MS MeCN/ water/0.1%TFA of Mass Spectrometer Method.This is clear Clear stream pours in separately in EtOAc (50ml), and uses saturated NaHCO₃Solution (50ml) washs, and is dried (MgSO₄) and be concentrated in vacuo, To title compound；19F NMR (400MHz, DMSO-d6): peak 1 is at-62ppm, and peak 2 is at-64.6ppm

Intermediate G:3-amino-5,6-two (trifluoromethyl) pyrazine-2-formylhydrazine

3-amino-5,6-two (trifluoromethyl) pyrazine-2-Ethyl formate (intermediate G1) (455mg, 1.501mmol) is existed Suspension in MeOH (10ml) monohydrate hydrazine (0.147ml, 3.00mmol) processes, and is stirred overnight under RT.Will reaction Mixture use water (50ml) dilutes, and with 1M HCl, pH is adjusted to pH4-5.Vacuum filters the yellow mercury oxide formed, and washes with water And air is dried, obtaining title compound, it the most directly uses；

LC-MS Rt=0.9min [M+H]+290.1 method 2minLC_v003.

1H NMR (400MHz, DMSO-d6): δ 10 (1H, s), 8.54 (2H, wide humps), 4.69 (2H, s).

Intermediate H

3-amino-6-bromo-5-(trifluoromethyl) pyridine-2-formylhydrazine

3-amino-6-bromo-5-trifluoromethylpyridin-2-methyl formate (intermediate A 4) (1.00g, 3.34mmol) is existed Suspension in anhydrous MeOH (20ml) (85 DEG C) under reflux stir 30 minutes, and subsequently with monohydrate hydrazine (324ul, 6.69mmol) process.This mixture is heated at reflux again 5 hours 30 minutes, and is cooled to RT.Add water, be collected by filtration Gained precipitates, and is dried in a vacuum furnace, obtains title compound, for buff white solid；

1H NMR(400MHz,DMSO-d6)δ9.50(1H,s),7.69(1H,s),7.19(2H,s),4.55(2H)。

LCMS:Rt=1.15min；[M+H]+299 method 2minLC_v002.

Intermediate compound I

3-amino-5-(trifluoromethyl) pyridine-2-formylhydrazine

Title compound is prepared, with 3-amino-5-trifluoromethylpyridin-2-formic acid according to the method similar with intermediate H Methyl ester (intermediate A 3) replaces 3-amino-6-bromo-5-trifluoromethylpyridin-2-methyl formate (intermediate A 4)；

LC-MS Rt=0.93min [M+H]+221.1 (method 2minLC_v002).

1H NMR(400MHz,DMSO-d6)δ9.80(1H,s),8.05(1H,s),7.51(1H,s),7.10(2H,s), 4.50(2H,s)。

Intermediate JR and JS

(R)-2-diazanyl-2-oxo-1-phenylethylcarbamate benzyl chloroformate (intermediate JR) and (S)-2-diazanyl-2-oxygen Generation-1-phenylethylcarbamate benzyl chloroformate (intermediate JS)

By (R)-2-(Benzyoxycarbonylamino)-2-ethyl phenylacetate (2.5g, 7.98mmol) in EtOH (20ml) Solution with monohydrate hydrazine (1.956ml, 39.9mmol) process, and under RT stir 6 days.Gained suspension is concentrated in vacuo, Obtain white solid.Use chiral separation to be purified according to following condition and obtain title compound:

Instrument: Gilson

Volume injected: 12ml

Flowing phase: normal heptane: EtOH60:40 (v/v)

Flow velocity: 60ml/min

Post: Chiralpak AS200x500mm, 20 μm

Detection UV:220nm

Intermediate JR:(R)-2-diazanyl-2-oxo-1-phenylethylcarbamate benzyl chloroformate

1H NMR(400MHz,DMSO-d6)d9.42(NH,s),7.94(NH,d),7.43(2H,d),7.35-7.25(8H, m),5.21(1H,d),5.03(2H,t),4.28(NH2,b s)

Intermediate JS:(S)-2-diazanyl-2-oxo-1-phenylethylcarbamate benzyl chloroformate

1H NMR(400MHz,DMSO-d6)d9.42(NH,s),7.94(NH,d),7.43(2H,d),7.35-7.25(8H, m),5.21(1H,d),5.07(2H,q),4.28(NH2,b s)

Although being appreciated that the specific embodiment party describing the present invention herein for illustrating purpose from foregoing description Case, but various change can be carried out without departing from the spirit and scope of the present invention.Therefore, the invention is not restricted to This, but be limited to the appended claims.

Embodiment:

Embodiment 1. compound of formula I or its officinal salt,

Wherein:

A is N or CR^4a；

X is

R²It is C₁-C₄Haloalkyl；

R³And R⁴Form oxo group (C=O) together；Or

M is 0,1,2 or 3；

R³⁰It is H or C₁-C₆Alkyl；

Embodiment 1.1: according to the compound of embodiment 1, wherein A is CR^4aAnd R^4aIt is selected from halogen, optionally by one Individual or multiple substituted C of halogen atom₁-C₄Alkyl；C₂-C₈Thiazolinyl and-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；Wherein-(C₀-C₄Alkane Base)-C₆-C₁₄Aryl can optionally be replaced by one or more Z substituent groups.

Embodiment 1.2: according to the compound of embodiment 1 or 1.1, wherein A is CR^4aAnd R^4aIt is selected from halogen, optionally The C replaced by one or more halogen atoms₁-C₄Alkyl；C₂-C₈Thiazolinyl and-(C₀-C₄Alkyl)-C₆-C₁₄Aryl.

Embodiment 1.3: according to the compound of embodiment 1,1.1 or 1.2, wherein A is CR^4aAnd R^4aIt is selected from chlorine, second Base, isopropyl, isopropenyl and phenyl；Wherein said phenyl can optionally be replaced by one or more Z substituent groups.

Embodiment 2. is according to the compound of embodiment 1,1.1,1.2 or 1.3, and wherein X is

The embodiment 3. compound as according to any one of embodiment 1 to embodiment 2, wherein

R¹It is selected from H；The C optionally replaced by one or more halogen atoms₁-C₈Alkyl；C₃-C₁₀Cycloalkyl；Optionally by one Individual or multiple substituted C of halogen atom₁-C₈Alkoxyl；Halogen；C₆-C₁₄Aryl；-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals, its Described in heterocyclic radical comprise at least one hetero atom selected from N, O and S；And NR¹¹R¹², wherein said aryl and heterocyclic radical are each Optionally replaced by one or more Z substituent groups.

The embodiment 4. compound as according to any one of embodiment 1-3, wherein

R¹It is selected from H, the C that optionally replaced by one or more halogen atoms₁-C₄Alkyl；C₃-C₁₀Cycloalkyl；Optionally by The substituted C of one or more halogen atoms₁-C₄Alkoxyl, and halogen.

The embodiment 5. compound as according to any one of embodiment 1-4, wherein

R¹It is selected from the C optionally replaced by one or more halogen atoms₁-C₄Alkyl；C₃-C₁₀Cycloalkyl；Optionally by one Individual or multiple substituted C of halogen atom₁-C₄Alkoxyl, and halogen.

Embodiment 6: the compound as according to any one of embodiment 1-4, wherein

The embodiment 7. compound as according to any one of embodiment 1-3, wherein

The embodiment 8. compound as according to any one of embodiment 1-7, wherein

R²It is CF₃。

Embodiment 9: the compound as according to any one of embodiment 1-8, wherein

Embodiment 10: the compound as according to any one of embodiment 1-9, wherein R⁴It is selected from H；Optionally by one Or the multiple substituted C of halogen atom₁-C₄Alkyl；The C optionally replaced by one or more halogen atoms₁-C₄Alkoxyl；- (CH₂)_m-NR¹⁷R¹⁸And OH；R¹⁷And R¹⁸It is H independently of one another；The C optionally replaced by one or more halogen atoms₁-C₈Alkyl.

Embodiment 11: the compound as according to any one of embodiment 1-10, wherein R⁵It is selected from H；Optionally by one Individual or multiple substituted C of halogen atom₁-C₄Alkyl；The C optionally replaced by one or more halogen atoms₁-C₄Alkoxyl； OH；CN；Halogen；-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；With-(C₀-C₄Alkyl)-3 to 14 yuan of heterocyclic radicals；Wherein said heterocyclic radical Comprising at least one hetero atom selected from N, O and S, wherein said aryl and heterocyclyl groups are the most one or more Z substituent group replaces.

Embodiment 12: the compound as according to any one of embodiment 1-11, wherein R³And R⁵It is connected with them Carbon atom forms 3-6 unit cycloalkyl together.

Embodiment 13: the compound as according to any one of embodiment 1-12, wherein R⁴And R⁵It is connected with them Carbon atom forms 5-6 unit cycloalkyl together or 5-6 unit comprises one or more heteroatomic heterocyclic radical selected from N, O and S, wherein Described heterocyclic radical is optionally replaced by one or more Z substituent groups.

Embodiment 14: the compound as according to any one of embodiment 1-8, wherein

R⁵It is selected from H；The C optionally replaced by one or more halogen atoms₁-C₄Alkyl；Optionally by one or more halogen The element substituted C of atom₁-C₄Alkoxyl；OH；CN；Halogen；-(C₀-C₄Alkyl)-C₆-C₁₄Aryl；With-(C₀-C₄Alkyl)-3 to 14 Unit's heterocyclic radical；Wherein said heterocyclic radical comprises at least one hetero atom selected from N, O and S, wherein said aryl and heterocyclic radical base Group is each optionally replaced by one or more Z substituent groups；Or

R⁴And R⁵Form 5-6 unit cycloalkyl together with the carbon atom being connected with them or 5-6 unit comprises one or more being selected from The heteroatomic heterocyclic radical of N, O and S, wherein said heterocyclic radical is optionally replaced by one or more Z substituent groups；

Embodiment 15: the compound as according to any one of embodiment 1-14, wherein

A is CR^4a；

X is

R²It is CF₃；

R³It is H, CH₃Or CF₃；

R⁴It is H or M_e；

R⁵It is phenyl ,-NR¹⁷R¹⁸Or OH；And

The embodiment 16. compound as according to any one of embodiment 1-15, wherein

A is CR^4a；

X is

R²It is CF₃；

R³It is H, CH₃Or CF₃；

R⁴It is H or Me；

R⁵It is-NR¹⁷R¹⁸Or OH；And

Embodiment 17: the compound as according to any one of embodiment 1-16, compound of formula I comprises Formula II compound Or its officinal salt:

Wherein A, R¹、R²、R³And R^4aAs defined in embodiment 1-16；And

R¹⁰¹It is selected from following group:

Embodiment 18: the compound as described in embodiment 17, wherein A is CR^4a, wherein R^4aIt is H.

Embodiment 19: the compound as described in embodiment 17 or 18, wherein A is CR^4a；

R¹It is the C optionally replaced by one or more halogen atoms₁-C₄Alkyl；

R¹⁰¹It is

Embodiment 20: the compound as described in embodiment 17, wherein R¹⁰¹It is

The embodiment 21. compound or pharmaceutically acceptable salt thereof as described in embodiment 1, is selected from:

2-(5-benzyl-[1,3,4] diazole-2-base)-5-trifluoromethylpyridin-3-base-amine；

(2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoropropyl- 2-alcohol (racemic)；

Embodiment 22: the compound as according to any one of embodiment 1-17, it is used as medicine.

Embodiment 23: the compound as according to any one of embodiment 1-17, it is used for treating inflammatory or obstructive Airway disorders or mucosal hydration.

Embodiment 24: the compound as according to any one of embodiment 1-17 is used for treating inflammatory or obstruction in preparation Purposes in the medicine of property airway disorders or mucosal hydration.

Embodiment 25: pharmaceutical composition, it comprises:

Compound as according to any one of embodiment 1-17 and

One or more pharmaceutically acceptable excipients.

Embodiment 26: pharmaceutical combination product, it comprises:

The first active component, including the compound as according to any one of embodiment 1-17；Become with the second activity Point, it is selected from osmotically active agent, ENaC blocker, anti-inflammatory agent, bronchodilator, antihistaminic, cough medicine, antibiotic and takes off Oxygen ribonuclease medicine, wherein the first and second active ingredient can be in identical or different pharmaceutical compositions.

Embodiment 27: prevent or treat disease or the method for disease that CFTR mediated, including:

At least one of effective dose is used as according to any one of embodiment 1-17 to the individuality needing this kind for the treatment of Compound.

Claims

1. compound of formula I or its officinal salt,

Wherein:

A is N or CR^4a；

X is

R¹It is selected from H；The C optionally replaced by one or more halogen atoms₁-C₄Alkyl；C₃-C₁₀Cycloalkyl；Optionally by one Or the multiple substituted C of halogen atom₁-C₄Alkoxyl；And halogen；

R²It is C₁-C₄Haloalkyl；

R⁴It is H；The C optionally replaced by one or more halogen atoms₁-C₈Alkyl；-(CH₂)_m-NR¹⁷R¹⁸；

R^4aIt is selected from H；The C optionally replaced by one or more halogen atoms₁-C₄Alkyl；C₂-C₈Thiazolinyl；-(C₀-C₄Alkyl)- C₆-C₁₄Aryl；Halogen；

R⁵It is the C optionally replaced by one or more halogen atoms₁-C₈Alkyl；-(CH₂)_m-NR¹⁷R¹⁸；-(CH₂)_m-OR⁴；-(C₀- C₄Alkyl)-C₆-C₁₄Aryl；Wherein said-(C₀-C₄Alkyl)-C₆-C₁₄Aryl is optionally replaced by one or more Z substituent groups； Or

R³And R⁴Form oxo group (C=O) together；

M is 0,1,2 or 3；

R¹⁷It is H；

R¹⁸It is H；

Z is halogen independently.

2. compound as claimed in claim 1, wherein X is

3. compound as claimed in claim 1, wherein R¹It is selected from the C optionally replaced by one or more halogen atoms₁-C₄ Alkyl；The C optionally replaced by one or more halogen atoms₁-C₄Alkoxyl, and halogen.

4. the compound as according to any one of claim 1-3, wherein

R²It is CF₃。

5. the compound as according to any one of claim 1-3, wherein

A is CR^4a；

X is

R¹It is selected from H；The C optionally replaced by one or more halogen atoms₁-C₄Alkoxyl；The most one or more The substituted C of halogen atom₁-C₄Alkoxyl；

R²It is CF₃；

R³It is H, CH₃Or CF₃；

R⁴It is H or Me；

R⁵It is phenyl ,-NR¹⁷R¹⁸Or OH；And

R¹⁷And R¹⁸It is H independently of one another.

6. the compound as according to any one of claim 1-3, wherein

A is CR^4a；

X is

R¹It is selected from the C optionally replaced by one or more halogen atoms₁-C₄Alkoxyl；Optionally by one or more halogen The element substituted C of atom₁-C₄Alkoxyl；

R²It is CF₃；

R³It is H, CH₃Or CF₃；

R⁴It is H or Me；

R⁵It is-NR¹⁷R¹⁸Or OH；And

R¹⁷And R¹⁸It is H independently of one another.

7. compound or pharmaceutically acceptable salt thereof as claimed in claim 1, is selected from:

2-(5-(3-amino-6-methoxyl group-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-three Fluorine propan-2-ol；

(R)-2-(5-(3-amino-6-methoxyl group-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1, 1-trifluoro propan-2-ol；

(S)-2-(5-(3-amino-6-methoxyl group-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1, 1-trifluoro propan-2-ol；

2-(5-(3-amino-5,6-two (trifluoromethyl) pyrazine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoropropyl- 2-alcohol；

(R)-2-(5-(3-amino-5,6-two (trifluoromethyl) pyrazine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoro Propan-2-ol；

(S)-2-(5-(3-amino-5,6-two (trifluoromethyl) pyrazine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoro Propan-2-ol；

2-(5-benzyl-[1,3,4] diazole-2-base)-5-trifluoromethylpyridin-3-base-amine；

2-(5-(3-amino-6-cyclopropyl-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-three Fluorine propan-2-ol；

2-(5-(3-amino-6-methyl-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoro Propan-2-ol；

2-(5-(3-amino-6-bromo-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoropropyl- 2-alcohol；

2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoro propan-2-ol；

(2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoropropyl-2- Alcohol；

(R)-2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoropropyl- 2-alcohol；

(S)-2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)-1,1,1-trifluoropropyl- 2-alcohol；

(R)-2-(5-(amino (phenyl) methyl)-1,3,4-diazole-2-base)-6-bromo-5-(trifluoromethyl) pyridine-3-amine； With

2-(5-(3-amino-5-(trifluoromethyl) pyridine-2-base)-1,3,4-thiadiazoles-2-base)-1,1,1-trifluoro propan-2-ol；

(R)-2-[5-(3-amino-4-chloro-6-methoxyl group-5-trifluoromethylpyridin-2-base)-[1,3,4] diazole-2-base]- 1,1,1-trifluoro-propyl-2-alcohol；

(S)-2-(5-(3-amino-4-chloro-6-methoxyl group-5-(trifluoromethyl) pyridine-2-base)-1,3,4-diazole-2-base)- 1,1,1-trifluoro propan-2-ol；

(S)-2-[5-(3-amino-4-ethyl-6-methoxyl group-5-trifluoromethylpyridin-2-base)-[1,3,4] diazole-2- Base]-1,1,1-trifluoro-propyl-2-alcohol；

(S)-2-[5-(3-amino-4-isopropenyl-6-methoxyl group-5-trifluoromethylpyridin-2-base)-[1,3,4] diazole- 2-yl]-1,1,1-trifluoro-propyl-2-alcohol；

(S)-2-[5-(3-amino-4-isopropyl-6-methoxyl group-5-trifluoromethylpyridin-2-base)-[1,3,4] diazole-2- Base]-1,1,1-trifluoro-propyl-2-alcohol；With

(S)-2-[5-(3-amino-6-methoxyl group-4-phenyl-5-trifluoromethylpyridin-2-base)-[1,3,4] diazole-2- Base]-1,1,1-trifluoro-propyl-2-alcohol.

8. the compound as according to any one of claim 1-7 is used for treating inflammatory or obstructive airway diseases or obtaining in preparation Purposes in the medicine of the disease benefiting mucosal hydration.

9. pharmaceutical composition, it comprises:

Compound as according to any one of claim 1-7, and

One or more pharmaceutically acceptable excipients.

10. pharmaceutical combination product, it comprises:

The first active component, including the compound as according to any one of claim 1-7；Second active ingredient, its choosing From osmotically active agent, ENaC blocker, anti-inflammatory agent, bronchodilator, antihistaminic, cough medicine, antibiotic and deoxyribose Nuclease medicine, wherein the first and second active ingredient are in identical or different pharmaceutical composition.